Acteoside promotes autophagy and apoptosis of hepatoma cells by regulating JNK signaling pathway / 中国中药杂志

This study explored the molecular mechanism of acteoside against hepatoma 22(H22) tumor in mice through c-Jun N-terminal kinase(JNK) signaling pathway. H22 cells were subcutaneously inoculated in 50 male BALB/c mice, and then the model mice were classified into model group, low-dose, medium-dose, and high-dose acteoside groups, and cisplatin group. The administration lasted 2 weeks for each group(5 consecutive days/week). The general conditions of mice in each group, such as mental status, diet intake, water intake, activity, and fur were observed. The body weight, tumor volume, tumor weight, and tumor-inhibiting rate were compared before and after administration. Morphological changes of liver cancer tissues were observed based on hematoxylin and eosin(HE) staining, and the expression of phosphorylated(p)-JNK, JNK, B-cell lymphoma-2(Bcl-2), Beclin-1, and light chain 3(LC3) in each tissue was detected by immunohistochemistry and Western blot. qRT-PCR was performed to detect the mRNA expression of JNK, Bcl-2, Beclin-1, and LC3. The general conditions of mice in model and low-dose acteoside groups were poor, while the general conditions of mice in the remaining three groups were improved. The body weight of mice in medium-dose acteoside group, high-dose acteoside group, and cisplatin group was smaller than that in model group(P<0.01). The tumor volume in model group was insignificantly different from that in low-dose acteoside group, and the volume in cisplatin group showed no significant difference from that in high-dose acteoside group. Tumor volume and weight in medium-dose and high-dose acteoside groups and cisplatin group were lower than those in the model group(P<0.001). The tumor-inhibiting rates were 10.72%, 40.32%, 53.79%, and 56.44% in the low-dose, medium-dose, and high-dose acteoside groups and cisplatin group, respectively. HE staining showed gradual decrease in the count of hepatoma cells and increasing sign of cell necrosis in the acteoside and cisplatin groups, and the necrosis was particularly obvious in the high-dose acteoside group and cisplatin group. Immunohistochemical results suggested that the expression of Beclin-1, LC3, p-JNK, and JNK was up-regulated in acteoside and cisplatin groups(P<0.05). The results of immunohistochemistry, Western blot, and qRT-PCR indicated that the expression of Bcl-2 was down-regulated in the medium-dose and high-dose acteoside groups and cisplatin group(P<0.01). Western blot showed that the expression of Beclin-1, LC3, and p-JNK was up-regulated in acteoside and cisplatin groups(P<0.01), and there was no difference in the expression of JNK among groups. qRT-PCR results showed that the levels of Beclin-1 and LC3 mRNA were up-regulated in the acteoside and cisplatin groups(P<0.05), and the level of JNK mRNA was up-regulated in medium-dose and high-dose acteoside groups and cisplatin group(P<0.001). Acteoside promotes apoptosis and autophagy of H22 cells in mice hepatoma cells by up-regulating the JNK signaling pathway, thus inhibiting tumor growth.

The In Vivo Intervention and Relative Mechanism of Genistein on the Inflammation and Thrombophilia in Lymphoma-Bearing Mice / 中国实验血液学杂志

OBJECTIVE@#To investigate the in vivo intervention and relative mechanism of Genistein (GEN) on tumor-associated inflammatory and tumor thrombophilia in lymphoma-bearing mice.@*METHODS@#Forty female Balb/c mice aged 5-6 weeks were injected with murine-derived Pro B-cell lymphoma cell line 38B9 to establish a lymphoma mouse model, which was randomly divided into control group, tumor-bearing group, GEN drug intervention group and cyclophosphamide (CTX）drug intervention group. Histopathologic was used to evaluate the tumorigenesis. Tumor formation was observed, and tumor tissues were collected of HE and immunohistochemical staining. ELISA and flow cytometry were used to detect the expression of inflammatory factors and the changes of thrombus indices in plasma after intervention of GEN and Cyclophosphamide (CTX) respectively. Immunohistochemistry method was used to detect the expression of CD19 in tomor tissues of tummor bearing mice.@*RESULTS@#After 14 days of tumor bearing, the mice were tumorigenic. The lymphoma cells were diffusely distributed in the tumor tissue and the expression of CD19 in the tumor tissue was positive. The inflammatory factors such as IL-6, NETs and CLEC-2, and thrombotic indices such as TF, FIB and D-D in lymphoma-bearing mice were significantly higher than those before tumor-injection and lower than those after drug-intervention (all P<0.05). The levels of CLEC-2 and D-D in GEN group were significantly lower than those in CTX group (P<0.05).@*CONCLUSION@#Tumor-associated inflammation and thrombophilia exist in lymphoma-bearing mice. GEN shows better anti-inflammatory and anti-thrombotic effects compared with CTX by interfering with tumor inflammatory factors.

Effects of increasing efficacy and decreasing toxicity of ginkgo flavone aglycone on doxorubicin in the treatment of liver cancer / 中国药房

OBJECTIVE To study the effects of increasing efficacy and decreasing toxicity of ginkgo flavone aglycone （GA） on doxorubicin （DOX）in the treatment of liver cancer. METHODS A tumor bearing model was established by inoculating liver cancer cell H 22 into the right axillary skin of ICR mice. The successfully modeled mice were randomly divided into model control group，DOX group （2.5 mg/kg，once every other day ，via tail vein ），GA group （30 mg/kg，once a day ，gavage）and GA+DOX group（the usage was the same as single drug groups ），with 6 mice in each group. The administration cycle was 15 days. The general growth of mice in each group were observed ，body weight and tumor weight were measured ，and the inhibition rate of tumor was calculated. Jin’s formula was used to evaluate the effect of combined medication （Q）. The serum level of alpha-fetal protein（AFP），the pathological changes of tumor tissue ，cell apoptosis and the expression of platelet-endothelial cell adhesion molecule-1（CD31）were detected in each group. The cardiac index，serum levels of B-type natriuretic peptide （BNP）and N-terminal pro-brain natriuretic peptide （NT-pro BNP ），pathological changes of heart and myocardial fibrosis degree were also detected. RESULTS The percentage of body weight change （except for GA group ） and tumor weights of DOX group，GA group and GA + DOX group were all decreased significantly，compared with model control group （P＜0.05 or P＜0.01），while tumor weight of GA+DOX gro up was significantly lower than DOX group （P＜0.01）. Inhibitory rates of tumor in 3 administration groups were 54.29％，42.50％ and 89.29％ respectively，and Q of two-drug combination was 1.21. The tumor tissues of mice in each administration group were necrotic to varying degrees ；the serum level of AFP and the expression of CD31 in tumor tissue were decreased significantly ，compared with model control group （P＜0.05 or P＜0.01）；the percentage of necrosis area of tumor tissue and the positive rate of apoptosis （except for single drug groups ）were significantly increased （P＜0.05 or P＜0.01），while positive rate of apoptosis in GA+DOX group was significantly higher than DOX group （P＜0.05）. Cardiac index of mice in DOX group was significantly lower than model control group （P＜0.01）；serum levels of BNP and NT-pro BNP in DOX group and GA+ DOX group were significantly higher than model control group （P＜0.05 or P＜0.01）；pathological changes of heart and the degree of myocardial fibrosis in GA+DOX group were lower than DOX group. CONCLUSIONS GA combined with DOX show synergistic antitumor effect. GA can strengthen the apoptosis promoting effect of DOX ，and can help to reduce the cardiotoxicity of DOX.

Study on the Activity of Lycorine to the in vivo Apoptosis of Tumor Cells in H 22-bearing Mice and Its Mechanism / 中国药房

OBJECTIVE：To investigate the activity of lycorine to the in vivo apoptosis of tumor cells in H 22-bearing mice and its mechanism. METHODS ：Kunming mice were inoculated subcutaneously with ascites of H 22 hepatoma mice in the armpit of forelimb to establish solid tumor model. After modeling ，mice were randomly divided into negative control group ，positive control group（hydroxycamptothecin 6 mg/kg），lycorine low-dose ，medium-dose and high-dose groups （10，20，40 mg/kg），with 10 mice in each group. Negative control group was given constant volume of normal saline intragastrically ，and administration groups were given relevant medicine intragastrically ，once a day ，for consecutive 7 days. After last medication ，the weight of tumor was detected and anti-tumor rate was calculated. Ascites tumor model of mice was established by intraperitoneal injection of H 22 hepatoma mice ascites ，and then were grouped with same method and given relevant medicine as above. After last medication ， survival time of mice was recorded and the life prolongation rate was calculated. The early apoptotic rate of tumor cells in mice was detected by flow cytometry. On the basis of normal control group （normal mice without tumor ），the mitochondrial membrane permeability of tumor cells in each group was investigated by Calcein AM staining. The changes of mitochondrial potential were investigated by Rhodamine 123 staining. Colorimetry and Western blot assay were adopted to detect the Caspase-3 activity and expression of apoptosis-related protein （Bcl-2，Bax，Cyt-C and Caspase- 9）. RESULTS ：Compared with negative control UN- group，the tumor weight of positive control group and lycorine PYSCT-2017208） groups were decreased significantly ，while the survival time was significantly prolonged ，and the early apoptotic rate of tumor cells was significantly increased （P＜0.05 or P＜0.01）；the anti-tumor rates were 39.41％ ， 23.36％ ， 36.50％ ， 56.93％，and life prolonga tion rates were 49.23％，29.09％， E-mail：ym913@yahoo.com.cn 50.19％，69.08％. Compared with normal control group ，the mitochondrial membrane permeability ，Caspase-3 protein activity and protein expression of Cyt-C and Caspase- 9 were significantly increased，while the mitochondrial membrane potential and Bcl- 2/Bax ratio were decreased significantly （P＜0.05 or P＜0.01）. Compared with negative control group ，mitochondrial membrane permeability and Bcl- 2/Bax ratio were decreased significantly in administration groups ，while mitochondrial permeability ，Caspase-3 protein activity and protein expression of Cyt-C and Caspase- 9 were significantly increased （P＜0.05 or P＜0.01）. CONCLUSIONS ：Lycorine can induce the apoptosis of tumor cells in H22-bearing mice ，the effects of which may be associated with opening mitochondrial membrane permeability transition pore to increase mitochondrial permeability ， decreasing mitochondrial membrane potential and up-regulating the expression of apoptosis-related proteins.

Exploration of Mechanism of Guben Qingyuan Prescription Combined with Androgen Deprivation Therapy Against Castration-resistant Prostate Cancer from AR/AR-V7 Signaling Pathway / 中国实验方剂学杂志

Objective:To explore the efficacy and mechanism of Guben Qingyuan prescription combined with androgen deprivation therapy （ADT） in the treatment of castration-resistant prostate cancer （CRPC）. Method:A CRPC-bearing mouse model was established. When the tumor volume reached about 100 mm³， 50 CRPC-bearing BALB/c nude mice were randomly divided into the model group， ADT group， and ADT+low-， medium-， high-dose Guben Qingyuan prescription groups， with 10 mice in each group. After grouping， it was ensured that there was no statistically significant difference in tumor volume between groups. The mice in the model group was treated with the same amount of normal saline （10 mL·kg^-1） by gavage， twice a day， while those in the other groups were provided with bicalutamide （5 mg·kg^-1） for intragastric administration， once a day， and then with goserelin （0.36 mg·kg^-1） for intraperitoneal injection on the 10th day. On the basis of ADT， the ones in the ADT+Guben Qingyuan prescription groups further received Guben Qingyuan prescription at the low （2.5 g·kg^-1）， medium （25 g·kg^-1）， and high doses （50 g·kg^-1） by gavage， twice a day. After 25 days of continuous administration， the tumor tissue was harvested for recording the tumor growth and calculating the tumor inhibition rate. The mRNA and protein expression levels of androgen receptor （AR）， androgen receptor splice variant-7 （AR-V7）， and prostate-specific antigen （PSA） were detected by real-time polymerase chain reaction （Real-time PCR） and Western blot assay. Result:The tumor inhibition rates of the ADT+low-， medium-， and high-dose Guben Qingyuan prescription groups were 27.95%， 46.71%， and 44.46%， respectively， and the inhibition rates in the ADT+medium- and high-dose Guben Qingyuan prescription groups were significantly increased as compared with that in the ADT group （<italic>P</italic><0.05）. As revealed by comparison with the ADT group， Guben Qingyuan prescription at the medium and high doses significantly down-regulated the mRNA and protein expression levels of AR， AR-V7， and PSA （<italic>P</italic><0.05）. Conclusion:Guben Qingyuan prescription combined with ADT is efficient in controlling the tumor growth in CRPC-bearing mice， which is related to the regulation of AR/AR-V7 signaling pathway.

Study on Anti-tumor Effects of Periplaneta americana Polypeptide PAP- 2 on H 22 Tumor-bearing Mice / 中国药房

OBJECTIVE： To investigate anti-tumor effects of Periplaneta americana polypeptide PAP-2 on H22 tumor-bearing mice. METHODS： The mice tumor-bearing model was established by subcutaneous injection of ascites of H22 hepatocellular carcinoma mice via axilla. 70 mice were randomly divided into model group （normal saline）， 5-FU group （positive drug control， 20 mg/kg）， P. americana extract skimmed cream group （200 mg/kg， calculated by extract）， CⅡ-3 group （polypeptide isolated from skimmed cream as main active ingredient， 200 mg/kg， calculated by extract） and polypeptide PAP-2 high-dose， medium-dose and low-dose groups （isolated from CⅡ-3， 200， 100， 50 mg/kg， calculated by monomer）， with 10 mice in each group. The mice in the 5-FU group were given intraperitoneal injection once every other day， while the mice in the other groups were given intragastric administration once a day， the administration cycle was 10 d. After medication， the changes of tumor were observed and the organs （spleen， thymus and liver） index were measured. Histopathological changes of tumor tissue were observed after HE staining. The contents of VEGF， IL-1β and IL-4 in serum were determined by ELISA. RESULTS： Skimmed cream， CⅡ-3 and different doses of PAP-2 could inhibit the growth of tumor in tumor-bearing mice to different extent and increase organ index， and PAP-2 showed a dose-effect relationship. The tumor inhibition rate （38.95％） of PAP-2 high dose group was significantly higher than those of skimmed cream group and CⅡ-3 group （P＜0.05）， which was close to that （40.87％） of 5-FU group （P＞0.05）. Spleen index， thymus index and liver index of mice in PAP-2 high dose group were significantly those of model group and CⅡ-3 group （P＜0.05）； and the liver index of mice in PAP-2 high dose group was significantly higher than that of skimmed cream group （P＜0.05）. In addition， PAP-2 could decrease the serum contents of VEGF and IL-4， and increased serum content of IL-1β， with high dose group showed significant difference compared with model group （P＜0.05）； the serum content of IL-1β of mice in PAP-2 high dose group was significantly higher the that of skimmed cream group and CⅡ-3 group （P＜0.05）， serum contnet of IL-4 in PAP-2 high dose group was significantly lower the  that of skimmed cream group and CⅡ-3 group （P＜0.05）， but the serum content in which was significantly lower than that of skimmed cream group and CⅡ-3 group（P＜0.05）. CONCLU- SIONS： P. americana polypeptide PAP-2 it has a certern anti-tumor effects on H22 tumor-bearing mice， and its can increase the index of organs of H22 tumor-bearing mice， decrease the contents of VEGF and IL-4 in serum， increase the content of IL-1β in serum.

Effect of Modified Lichongtang Combined with 5-Fluorouracil on Epithelial Mesenchymal Transition in H22 Tumor-bearing Mice / 中国实验方剂学杂志

Objective: To investigate the effect of modified Lichongtang combined with 5-fluorouracil (5-FU) on tumor epithelial-mesenchymal transition (EMT) in H22 tumor-bearing mice. Method: Mouse models of transplanted hepatoma were constructed. After tumor formation, they were randomly divided into 4 groups:blank group, 5-FU group(2.5 mg·kg-1 5-FU intraperitoneal injection), modified Lichongtang combined with 5-FU group (5-FU+Chinese medicine group), and modified Lichongtang group (Chinese medicine group,25 g·kg-1 gavage),n=10 in each group. The effect of modified Lichongtang combined with 5-FU on the tumor inhibition rate of subcutaneous transplanted tumor was observed. The gene expression levels of E-cadherin,N-cadherin,Snail,and Twist in transplanted tumor were observed by Real-time PCR. The protein expression levels of E-cadherin,N-cadherin,Snail,and Vimentin were detected by using Western blot. Result: The tumor inhibiting rate was 59.18%,84.42%,and 10.39% respectively in 5-FU group, 5-FU+Chinese medicine group,and Chinese medicine group. All of these can inhibit the growth of liver cancer transplantation tumor, and the tumor inhibiting rate of 5-FU+Chinese medicine group was significantly higher than that in 5-FU group (PPPPPPPPPPPConclusion: Modified Lichongtang, 5-FU and their combination have inhibitory effect on the growth of transplanted tumors of hepatocarcinoma mice, and the combination of the two drugs can enhance the effect of chemotherapy and to some extent inhibit the toxicity of 5-FU. The mechanism may be related to the inhibition of liver cancer EMT.

Effect on tumor growth inhibition activity of Gekkoswinhonis Guenther with different extent of broiling on H22 tumor-bearing mice / 药学学报

This study was designed to investigate the effect on tumor growth inhibition activity of lizards (Gekkoswinhonis Guenther) with different extent of broiling. Samples were prepared by a traditional drying method combined with broiling on clay tiles. Four groups of samples were all dried before broiling. Group A was without broiling; group B was mildly broiled; group C was moderately broiled; and group D was heavily broiled. Crispiness was detected by the sizes of the generated fragments of different groups and crispiness increased with broiling. Sensory evaluation of vision and olfaction was performed, and scores were generated by evaluators. Moderately broiled group had the highest total score in sensory evaluation. Water content and content of water-soluble extracts were detected according to Chinese Pharmacopoeia. With the increasing broiling extent, content of water-soluble extracts increased while water content decreased. Soluble protein concentration was detected by bicinchoninic acid (BCA) kit and sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) with the same crude drug content. Soluble protein concentration decreased with the increasing broiling extent. With equal loading of proteins at the same concentration, soluble protein diversity was detected by SDS-PAGE. Band difference was marked by red boxes. Soluble protein molecule weights showed significant difference with the increasing broiling extent. H22 tumor-bearing mice model was established and used to detect tumor growth inhibition rate and immune organ index. Life quality of mice was evaluated. Mice treated with Gekkoswinhonis Guenther had better appetites and higher average weights compared with positive control group treated with fluorouracil (5-FU). Animal experiments were approved by the Ethics Committee of Beijing University of Chinese Medicine. Group A had the highest tumor growth inhibition rate (34.11%), followed by Group B (29.14%) and Group D (28.43%), Group C (21.98%) had the lowest tumor growth inhibition rate, but sensory evaluation was on the contrary. These results indicated that moderately broiling improved sensory evaluation but reduced the tumor growth inhibition activity of Gekkoswinhonis Guenther. The best tumor growth inhibition activity appeared when water content was 7.71%.

An-tumor activity of sesquiterpenoids from ginseng in S180 tumor-bearing mice and its mechanism / 吉林大学学报(医学版)

Objective: To investigate the anti-tumor activity in vivo of sesquiterpenoids from ginseng (SPG) in the S180 tumor-bearing mice, and to clarify its mechanism Methods: The ICR male mice were used to establish the S180 tumor-bearing models. The model mice were randomly divided into model group, 5-fluorouracil (5-FU) group (20 mg • k g- 1 , ip), low dose of SPG group (2. 5 mg • k g- 1 , ig), and high dose of SPG group (10. 0 mg • k g- 1 , ig), 8 mice in each group; another 8 mice were selected and used as blank control group. Fourteen days after administration, the blood samples were collected from the eyes of all mice, and they were sacrificed by cervical dislocation, then the tumor tissue was excised and weighed; the inhibitory rate of tumor was calculated; the serum levels of alanine aminotransferase (AST), aspartate aminotransferase (ALT), urea nitrogen (BUN), interleukin-2 (IL-2), tumor necrosis factor-a (TNF-a) and vascular endothelial growth factor (VEGF) of the mice in various groups were measured; the pathological changes of the tumor tissue of the mice in various groups were observed by HE staining; the expression levels of anti-apoptotic factor Bel-2, VEGF, p38 against mitogen-activated protein kinase (p38MAPK) and p-p38 against mitogen-activated protein kinase (p-p38MAPK) in the tumor tissue of the mice in various groups were analyzed by Western blotting method. Results: The inhibitory rates of the mice in SPG groups were increased significantly as the increase of SPG dose, and the inhibitory rate of tumor of the mice in high dose of SPG group was 76. 29%. Compared with model group, the serum levels of IL-2 and TNF-a of the mice in SPG groups were significantly increased (P < 0 . 01) and the levels of ALT, AST, BUN and VEGF were significantly decreased (P < 0. 05 or P< 0. 01). The HE staining results showed that the cells in model group were arranged neatly, a large number of nuclei were observed and the growth state was good; in low and high doses of SPG group, the number of nuclei in the tumor tissue of the mice was significantly reduced, the arrangement was loose, and a large area of necrosis occurred. The Western blotting results showed that compared with model group, the expression levels of Bcl-2 and VEGF proteins were significantly decreased (P < 0 . 01) and the expression levels of p-p38MAPK protein in low and high doses of SPG groups were significantly increased (P < 0.01). Conclusion: SPG has a good anti-tumor effect in the S180 tumor-bearing mice, and its mechanism may be associated with the decreasing of Bcl-2 and VEGF expressions and activation of p38 MAPK protein channel.

Anti-tumor and immune-modulating effect of decoction in mice bearing hepatoma H22 tumor / 南方医科大学学报

OBJECTIVE@#To investigate the antitumor activity of decoction and study its liver and kidney toxicity and its effect on the immune system in a tumor-bearing mouse model.@*METHODS@#Hepatoma H22 tumor-bearing mouse models were randomized into model group, cyclophosphamide (CTX) group, and low-, moderate-, and high-dose decoction groups (JW-L, JW-M, and JW-H groups, respectively). The antitumor activity of decoction was assessed by calculating the tumor inhibition rate and pathological observation of the tumor tissues. Immunohistochemistry was used to detect the expressions of Bax, Bcl-2, Bax/Bcl-2 and caspase-3 in the tumors. The liver and kidney toxicity of decoction was analyzed by evaluating the biochemical indicators of liver and kidney functions. The immune function of the tumor-bearing mice were assessed by calculating the immune organ index, testing peripheral blood routines, and detection of serum IL-2 and TNF-α levels using enzyme-linked immunosorbent assay.@*RESULTS@#Compared with that in the model group, the tumor mass in CTX, JW-M and JW-H groups were all significantly reduced ( < 0.05) with cell rupture and necrosis in the tumors. Immunohistochemistry revealed obviously up-regulated expressions of Bax and caspase-3 and down- regulated expression of Bcl-2 protein with an increased Bax/Bcl-2 ratio in CTX, JW-M and JW-H groups. Treatment with decoction significantly reduced Cr, BUN, AST and ALT levels, improved the immune organ index, increased peripheral blood leukocytes, erythrocytes and hemoglobin levels, and up-regulated the levels of TNF-α and IL-2 in the tumor-bearing mice. These changes were especially significant in JW-H group when compared with the parameters in the model group ( < 0.01).@*CONCLUSIONS@# decoction has a strong anti-tumor activity and can improve the liver and kidney functions of tumor-bearing mice. Its anti-tumor effect may be attributed to the up-regulation of Bax, caspase-3, TNF-α and IL-2 levels and the down-regulation of Bcl-2 expression as well as the enhancement of the non-specific immune function.

Suppression effect of secreting-trail adenovirus on the prostate cancer PC-3 cells in the tumor bearing mice / 实用医学杂志

Objective To investigate the suppression effect of secreting-trail adenovirus on the prostate cancer PC-3 cells in the tumor bearing mice. Methods Apoptosis of PC-3 cells was measured by DAPI staining in vitro . Invasion of PC-3 cells was measured by transwell chamber assay. The tumor bearing mice were prepared and used to test the inhibition rates in different groups. The protein expression of TRAIL in different groups tumor bearing mice was determined by immunohistochemistry. The variation of cell apoptosis in different groups of tumor bearing mice was detected by TUNEL assay. Results DAPI staining showed that the apoptosis level in Ad-sTRAIL group was significantly higher than that in the control group and that in the Ad-LacZ group ,respectively. The number of PC-3 cells invading the inferior chamber in the Ad-sTRAIL group(24.8 ± 3.70) was significantly decreased compared with that in the control group(47.6 ± 4.28,q=10.68,P<0.01)and that in the Ad-LacZ group (49.6 ± 4.39,q=11.61,P<0.01). Growth inhibition assays in vivo showed the inhibition rate of Ad-sTRAIL was 3 d 8.15%,6 d 11.55%,9 d 17.23%,12 d 20.05%,15 d 27.18%,18 d 34.27%,and 21 d 33.08%,respective-ly. Immunohistochemistry results showed that the expression level of TRAIL in tumors of Ad-sTRAIL group tumor-bearing mice was significantly higher than that in the control group and that in the Ad-LacZ group. TUNEL assay showed that the apoptosis level of tumor bearing mice tumor cells in the Ad-sTRAIL group was significantly higher than that in the control group and that in the Ad-LacZ group. Conclusion The secreting-trail adenoviral vector could inhibit the growth of tumor bearing mice tumor cells,furthermore it could induce the apoptosis of PC-3 cells in the tumor bearing mice.

Roles of human leukocyte antigen-G ( HLA-G) in MDSC proliferation, M1/M2 macrophage differ-entiation and tumor immune evasion / 中华微生物学和免疫学杂志

Objective To investigate the roles of human leukocyte antigen-G ( HLA-G) in mye-loid-derived suppressor cell (MDSC) proliferation and M1/M2 macrophage differentiation in C57BL/6-NCI-H446-G tumor-bearing mice for better understanding the mechanisms of HLA-G involved in tumor immune evasion. Methods NCI-H446 ( human small cell lung cancer cells) and NCI-H446-G ( NCI-H446 cells ex-pressing HLA-G) cells were labeled with CFSE at a final concentration of 1μmol/L. CFSE fluorescence lev-els were measured by flow cytometry at different time points. Mouse tumor models were established by subcu-taneous injection of C57BL/6 mice with NCI-H446 and NCI-H446-G cells, respectively. PBS was used to set up negative control group. The mice in each group were sacrificed to collect tissue samples on 5 d, 10 d, 15 d and 20 d after injection. The percentages of splenic CD11b+Gr1+MDSCs, F4/80+CD80+M1 and F4/80+CD206+M2 macrophages were analyzed by flow cytometry. Results Steady expression of HLA-G in NCI-H446-G cells was confirmed by Western blot and flow cytometry. HLA-G enhanced the proliferation of NCI-H446 cells. Tumor size increased dramatically in tumor-bearing mice in the first five days and then de-creased over time. The tumor-bearing mice in the NCI-H446-G group had larger tumor than those in the NCI-H446 group in every time point (P<0. 05) and required longer time to fully reject the tumor. Compared with the PBS and NCI-H446 groups, the percentage of splenic MDSCs in tumor-bearing mice was significantly in-creased in the NCI-H446-G group (P<0. 05). Moreover, the ratio of M1/M2 in NCI-H446-G tumor-bearing mice was much lower than that in the other two groups (P<0. 05). Conclusion This study indicated that HLA-G could increase the percentage of MDSCs and decrease the ratio of M1/M2, which might illustrate the role of HLA-G in tumor immune evasion and its potential clinical significance in cancer immunotherapy.

A Research on the Anti-tumor Effects of Acid-Soluble Polysaccharides from Gloeostereum incamatum in H22 Tumor Bearing Mice / 世界科学技术-中医药现代化

This study aimed at exploring the inhibitory effect behind its mechanism on acid-soluble polysaccharides from G.incamatum in transplanted H22 tumor mice.Different indices,including tumor inhibitory rate,organ index of liver,thymus and spleen,IL-2,IFN-γ and TNF-α were detected for the evaluation of anti-tumor effects and the mechanism.Furthermore,HE staining and TUNEL assay were adopted to investigate the pathological changes of tumor tissue and cell apoptosis,respectively.As a result,the three dose groups of acidsoluble polysaccharides of G.incamatum successfully inhibited the proliferation of tumor cells,while organ indexes of spleen and thymus were improved and serum IL-2,IFN-γ and TNF-α increased.H&E staining and TUNEL assay showed the polysaccharides induced cell apoptosis,playing a significant role in the inhibition of tumor growth.In conclusion,acid-soluble polysaccharides of G.incamatum possessed significant anti-tumor effects,behind which the mechanism could be related to the regulation of immune regulation,cell apoptosis,and the protection of liver function.

Effects of Dangguibeimukushen pill on expressions of MMP13 and bFGF on H22 hepatocarcinoma xenograft mice / 国际检验医学杂志

Objective To study the effects of supplemental Dangguibeimukushen pill prescription on the expressions of MMP13 and bFGF in tumor tissues on H22 hepatoma bearing mice.Methods H22 hepatoma bearing mice was employed to carry out anti-tumor experiment in vivo in this study.A total of 60 mice were randomly divided into model group,DDP positive control group,low and high dosage of supplemental Dangguibeimukushen pill treatment group,supplemental Dangguibeimukushen pill combined with DDP group.In each gruop,gastric infusion was performed continuously 14 d.The tumor growth and the general conditions of mice were recorded.After the last administration of gavages orally treatment,all mice were anaesthetized and killed by cervical disloca-tion method to obtain completely tumor tissue for further HE staining measure and detection of MMP13 and bFGF in tumor tissue with the method of RT-qPCR and immunohistochemistry.Results The tumor volume of model group and low dosage of supple-mental Dangguibeimukushen pill treatment group were larger than that of other groups,futhermore,ulceration were found inside tumor,and basic integrity were observed of tumor capsule.However,in all the treatment groups,the tumor volume was relatively smaller,and the capsule of tumor was incomplete,ulceration of different degrees and bleeding when tumor was peeled off.In combi-nation groups,the tumor volume was much smaller,and the tumor had incomplete capsule and less ulceration inside.Pathological observation showed that connective tissue was found inside tumor,cells were sparsely spread and the number of necrosis cancer cells were increased in all treatment groups.RT-qPCR detection showed that the mRNA expression of MMP13 and bFGF in each treat-ment group were decreased comparing with the model group,and the difference was statistically significant(P <0.05).Immunohis-tochemical detection showed that the expression of MMP13 and bFGF in the combined group were less colored and weakly positive expression comparing with the positive group or the pure Chinese medicine treatment group,the difference was statistically signifi-cant(P <0.05).Conclusion Dangguibeimukushen pill could downregulate the expression of MMP13 and bFGF of tumor tissue in H22 hepatocarcinoma xenograft mice at mRNA and protein levels,and played a vital role in inhibiting cancer invasion and angiogen-esis,reducing toxics and side effects and enchancing its efficiency.

The in vitro and in vivo antiangiogenenic effect of galangin / 国际药学研究杂志

Objective To investigate the antiangiogenic effect of galangin in vitro and in vivo.Methods The inhibitory ef?fect of galangin on human umbilical vein endothelial cells(EaHy926)was tested by sulphorhodamine(SRB)method,the EaHy926 endothelial cell migration was assessed using in vitro model system,and the in vivo antiangiogenic effect of galangin at 1,5 and 10 μg was evaluated using the chorioallantoic membrane(CAM)model.The H22 tumor-bearing model was established using BALB/c nude mice,which were divided into five groups:The model group,positive control group〔ip cyclophosphamide 20 mg/(kg·d)〕and the galangin 10,20 and 40 mg/(kg·d)groups,respectively.After the daily administration for 15 consecutive days,the tumors grown in the nude mice were examined and the microvessel density(MVD)was tested via examining the expression of CD31 in the tumor tissues by immunohistochemistry in order to evaluate the effect of galangin to the tumor growth and the angiogenesis of the tumors. Result Galangin inhibited both the growth,migration and Matrigal tubule of EaHy926 cells in a dose-dependent manner in the in vitro test. Further in the in vivo test,galangin could also obviously inhibit the angiogenesis of CAM at the 5 and 10 μg concentration.The tumor mass and the relative tumor volume in the 20 and 40 mg/(kg·d)galangin groups and the positive control cyclophosphamide group were significantly lower(P<0.05)than those in the model group in the in vivo nude mice test. The tumor inhibitory rates of those three groups were 34.17%,79.73% and 55.75%,respectively.The MVD was also significantly lower in the high dosage galangin groups than that in the model group. Conclusion Galangin showed the obvious anti-angiogenenic effect both in vitro and in vivo in the present study.

Anti-tumor Effect of Ginsenoside Rg1 Pyrolysis Products (HPPRg1) on H22 Tumor-bearing Mice / 中国药学杂志

OBJECTIVE: To investigate the antitumor effect and molecular mechanism of ginsenoside Rg1 pyrolysis products (HPPRg1) on H22 tumor bearing mice. METHODS: To establish tumor model of transplanting H22 tumor-bearing mice and observe the anti-tumor effects of HPPRg1, H22 tumor-bearing mice were randomly divided into groups of control, model, cyclophosphamide (CTX, 30 mg·kg-1), low dosage of HPPRg1 (HPPRg1-L, 10 mg·kg-1), middle dosage of HPPRg1 (HPPRg1-H, 20 mg·kg-1) and high dosage of HPPRg1 (HPPRg1-H, 40 mg·kg-1) groups, respectively. Through evaluating inhibition rates of tumors, organ indices, and levels of TNF-α, IFN-γ and IL-2 to observe the anti-tumor effect of HPPRg1. In addition, H&E and Hoechst 33258 straining were used to observe the apoptosis of H22 tumor cell. RESULTS: Compared with the model group, the three dose groups of HPPRg1 can inhibit tumor proliferation. Mainly through the inhibition of tumor cell proliferation and pro-apoptosis to exert anti-tumor effect. CONCLUSION: HPPRg1 has a significantly inhibitory effect on H22 tumor-bearing mice, the mechanism may related to promote apoptosis of tumor cells and improve immunity.

Antitumor effect of alcohol extract of pileostegia tomentella on H22 tumor-bearing mice / 中国药学杂志

OBJECTIVE: To study the antitumor effect of Pileostegia tomentella 95% alcohol extract (PTAE) on H22 tumor-bearing mice and its possible mechanisms. METHODS: Sixty mice were chosen and mouse models bearing H22 solid tumor were established in fifty mice, and the others were as normal control. H22 tumor-bearing mice were randomly divided into five groups:model control group, fluorouracil group(20 mg·kg-1), PTAE high, middle and low-dose group(180, 90, 45 g·kg-1 of crude drug, respectively). The mice in treatment groups were intragastric administration respectively, meanwhile, the mice in normal control and model groups were treated with the same volume of distilled water, once a day for ten days. The blood was collected from eyeball in all mice, and the serum were separated and detected by ELISA for IL-2 and TNF-α. Then the mice were put to death. Their tumors, thymuses and spleens were separated and weighted, and the tumor inhibitory rates, thymus and spleen indexes were calculated. The pathological change of tumor tissue was observed. RESULTS: Compared with model control group, the tumor weights of PTAE high and middle-dose groups were significantly decreased (P<0.05, P<0.01), the tumor inhibitory rates were 37.44% and 38.46% respectively. The spleen index of PTAE middle-dose group was increased significantly (P<0.01). The level of IL-2 in serum of tumor-bearing mice in the PTAE high-dose group was increased significantly(P<0.05), and the level of TNF-α in serum (P<0.01) could be increased significantly in the PATE high, middle and low-dose groups. CONCLUSION: Pileostegia tomentella 95% alcohol extract has antitumor activity, its mechanism may be developed by immuno-regulation.

Development and evaluation antitumor activity of PEGylated liposomal doxorubicin on tumor-bearing BALB/c-Foxn1nu mice model.

Doxorubicin hydrochloride is an antitumor antibiotic derived from anthracyclines. It has had limited use because of its dose-related cardiotoxicity and myelosuppression. Liposomes have been used as a vehicle for administration of pharmaceutical drugs because of their ability to improve the delivery of drugs to tumors, increase therapeutic efficacy, and decrease toxicity to normal cells. The aim of this study is to prepare a new liposomal dxorubicin on a large-scale and evaluate its antitumor activity in vivo. Liposomes were formed using the hydration of a thin lipid film method, and doxorubicin was loaded through a pH gradient technique. Based on TEM images, large lamellar vesicles (LUV) were formed, with sizes of 95 ± 10 nm, having a polydispersity index of 0.138 ± 0.02 and zeta potentials of about -27.8 ± 2.15 mV. The entrapment efficiency was approximately 97%. The therapeutic activity of PEGylated liposomal doxorubicin formulations was studied on human colorectal carcinoma HT 29 tumor-bearing BALB/c-Foxn1nu mice models. Our results have shown that liposome preparation can reduce the tumor volume and increase the survival rate and survival time as compared with Lipo Dox. PEGylated liposomal doxorubicin demonstrated much stronger antitumor activities, and statistical differences were significant when compared with free doxorubicin.

Inhibitory Effect of Chenodeoxycholic Acid-verticinone Ester on Tumor Growth of H22-bearing Mice / 医药导报

Objective To evaluate the antitumor effects of chenodeoxycholic acid-verticinone ester ( CDCA-Ver ) on tumor growth and immune system of H22-bearing mice. Methods Antitumor activity against a solid tumor mass was evaluated in Kunming mice. H22 cells were transferred into the abdomen cavity of Kunming mice. H22 cells were inoculated through subcutaneous injection at the right armpit of the mouse to establish a solid tumor model. At 24 h after H22 tumor cells inoculation, 40 tumor-bearing Kunming mice were randomly divided into 4 groups according to random number table ( n=10 each group):model control group, cyclophosphamide ( CTX) group, intraperitoneal CDCA-Ver injection group and intravenous CDCA-Ver injection group. In model control group, sterile 0. 9% sodium chloride solution (10 mL·kg-1 ) was intraperitoneally injected once daily. In CTX group and intraperitoneal CDCA-Ver injection group, CTX (20 mg·kg-1 ) and CDCA-Ver (20 mg·kg-1 ) was intraperitoneally injected once daily, respectively. In intravenous CDCA-Ver injection group, CDCA-Ver ( 20 mg · kg-1 ) was injected through tail vein once daily. CDCA-Ver, CTX and NS were injected into the mice of the experimental groups once daily for 10 days, respectively. The dose volume was 0. 1 mL · ( 10 g )-1 body weight. The positive control drug was cyclophosphamide. Ten mice were treated with 20 mg · kg-1 CDCA-Ver through intravenous injection ( i. v. ) . Ten mice were treated with 20 mg·kg-1 CDCA-Ver through intraperitoneal injection. The thymus and spleen indices and the tumor inhibition rate were assessed, and histopathological examination with haematoxylin and eosin ( H&E) staining was carried out to evaluate the antitumor effects of CDCA-Ver. Results CDCA-Ver ( ivor ip) suppressed the growth of solid tumor in H22-bearing mice. The inhibition rate was 48. 3% at the dose of 20 mg·kg-1 CDCA-Ver (ip). There was no significant difference between CDCA-Ver (ip) and CTX treated group (P<0. 05). Compared with the control, the weight of thymus and spleen of CDCA-Ver (ip) treated group was not obviously changed. But a significant weight loss of thymus and spleen in CTX group was observed, which was attributed to the immune suppression from CTX. The thymus and spleen indices in the CTX-treated mice were significantly lower than those of the control group (P<0. 01). We further conducted histopathological examination to confirm the results. The immune system was not suppressed by CDCA-Ver ( ip ) in tumor-bearing animals. The low toxicity of CDCA-Ver was an outstanding advantage for the development of newly anticancer drug. Conclusion CDCA-Ver treatment can significantly inhibit tumor growth in mice.

Anti-breast cancer properties and toxicity of Dillenia suffruticosa root aqueous extract in BALB/c mice

Objective: To determine the anti-breast cancer activities and the safety oral consumption of Dillenia suffruticosa root aqueous extract (DRAE) in BALB/c mice. Methods: In the anti-breast cancer study, female BALB/c mice were divided into five groups (n = 12), which were (1) positive control (with breast cancer, untreated), (2) negative control (without breast cancer, untreated) and other three groups of mice with breast cancer treated with 1 000, 500 and 250 mg/kg of DRAE, respectively, by oral gavage for 28 days. All mice except from the negative control group were injected into the mammary fat pad with 4T1 cells (1 × 105 4T1 cells/0.1 mL of phosphate buffer solution). DRAE was administered orally on Day 11 after the tumor has developed. Results: The tumor volume of the 1 000 mg/kg of DRAE group reduced significantly compared to the positive control while treatment with 500 mg/kg of DRAE had signif-icantly inhibited metastasis to the heart. In the acute toxicity study, treatment with up to 5 000 mg/kg of DRAE was not toxic to the animals, indicating its safety when a large amount of this plant extract was ingested. Based on the sub-acute toxicity study, treatment of the highest dose of DRAE (1 000 mg/kg) had mild liver toxicity indicated by mild focal hemorrhage. Conclusions: DRAE possesses anti-breast cancer properties but at the same time it shows mild toxicity to the liver. The non observable adverse effect dose for DRAE is 500 mg/kg.