Advances in research and application of breast cancer organoids / 中国组织工程研究

BACKGROUND: Breast cancer is one of the most common malignant tumors in women. Its incidence rate is increasing year by year and tends to be younger. It seriously threatens women’s health. Therefore, it is particularly important to establish an ideal breast cancer model that can accurately simulate the tumor in vivo. Organoid is a new three-dimensional cultural model in vitro, which recapitulates key aspects of in vivo tissue or organ. In recent years, researches based on organoids have covered many kinds of tumors. OBJECTIVE: To review the research progress and application of breast cancer organoids, in order to provide a new research way for personalized treatment of breast cancer. METHODS: Using the key words of “organoid, breast cancer organoids, cancer organoids, mammosphere, three-dimensional culture” in English and Chinese, respectively, the first author retrieved relevant articles published from January 1980 to February 2020 in CNKI, Wanfang, and PubMed databases. The type of the article was not limited. After removal of the articles that were not related to the purpose of the study or repetitive, 66 articles were finally analyzed. RESULTS AND CONCLUSION: This review introduced organoid technology briefly and retraced the process of exploring suitable culture conditions to establish breast-cancer-origin organoids. Also, we concluded latest development of its applications and research progress. Breast cancer organoids have a wide range of application prospects in disease modeling, tumor pathogenesis, drug screening and other aspects, which provide a reliable model for breast cancer research and treatment, and in particular, open up a new perspective for personalized treatment of breast cancer.

Application of modified closed biopsy in rabbit model of VX2 transplanted bone tumor / 解放军医学杂志

Objective To explore the application value of modified closed biopsy technique in puncture biopsy of rabbit model of VX2 transplanted bone tumor. Methods VX2 tumor tissue was cut from rabbit with VX2 tumor and transplanted into the bilateral tibia of 30 rabbits through the tibial plateau to make the model of VX2 transplanted bone tumor. Seven days after modeling, the proximal tibia puncture biopsy was performed under the guidance of X-ray, and the biopsy specimen was examined pathologically. The left leg was biopsied with modified closed biopsy technique (experimental group), and the right leg was biopsied with hollow needle (control group). On the 14th day after modeling, all rabbits were executed after X-ray examination around the puncture hole, and the soft tissue around the puncture hole was taken for pathological examination. Results By the end of the experiment, a total of 3 rabbits died, and finally 27 rabbits were included in the study. Tumor cells were detected in all the intramedullary specimens obtained by puncture biopsy. On the 14th day after modeling, X-ray examination showed that, compared with control group, the incidence of periosteal reaction and extraosseous high density shadow around the puncture hole, and the tumor cell metastasis rate were lower [14.81%(4/27) vs. 40.74%(11/27); 29.63%(8/27) vs. 100.00%(27/27)], the differences were statistically significant (P<0.05). Conclusions Both the modified closed biopsy technique and puncture needle aspiration biopsy can provide sufficient biopsy tissue for diagnosis of VX2 transplanted bone tumor in rabbits. Meanwhile, the improved closed biopsy technique can prevent local metastasis of tumor cells along the puncture channel to some extent.

Comparing tumor suspension injection and tissue explant to establish rabbit vx2 subcutaneous tumor model / 中国组织工程研究

BACKGROUND: There are many methods to establish VX2 subcutaneous tumor model, and the existing modeling methods have their own advantages and disadvantages. To meet the needs of tumor experiments on animals, the method of establishing an animal tumor model with high-quality and high-efficiency is necessary. OBJECTIVE: To explore more simple and efficient way of modeling, provide rabbit VX2 subcutaneous tumor experiment with large quantities of high-quality animal models by comparing different methods to establish rabbit VX2 subcutaneous tumor model. METHODS: Sixty-six male New Zealand white rabbits were enrolled. Two of the 66 rabbits were used to prepare VX2 tumor-bearing rabbits, and tumor tissues were taken from the tumor-bearing rabbits to prepare tumor tissue blocks and tumor tissue suspensions. There were two groups in the experiment. In tumor tissue suspension group (n=20), the rabbits were injected with 0.15 mL of tissue suspension on the medial side of bilateral hind limbs after anesthesia; in tumor tissue block group (n=20), tumor tissue blocks were implanted subcutaneously on the medial side of bilateral hind limbs after anesthesia. Two tumor-bearing rabbits from each group were subjected to the corresponding vaccination methods, each passed for five generations. Tumor inoculation time in the two groups was record and compared. The tumor size and growth were observed by ultrasound with 2-D and CDFI mode. Tumor-take rate and serial passage of tumor tissues were observed and compared between the two groups. The experimental protocol was approved by the Animal Experiment Ethics Committee of the Army Medical University of PLA (approval No. AMUWE2020016) RESULTS AND CONCLUSION: The tumor inoculation time in the tissue suspension group [(75.70±11.16) s] was significant shortened compared with that in the tissue block group [(100.80±9.21) s; P=0.00]. The tumor-take rate was significantly higher in the tissue suspension group than the tissue block group (95% vs. 60%; P < 0.05). The tumor size was significantly larger in the tissue suspension group than the tissue block group (P < 0.05). The rate of tumor tissue series passage was significantly higher in the tissue suspension group than the tissue block group (95% vs. 65%; P < 0.05). Therefore, tissue suspension method for making the model of rabbit VX2 subcutaneous tumor is simpler and more efficient compared with the tissue block method.

Establishment of Animal Models for Prediction of Therapeutic Efficacy of Molecularly Targeted Agents on Advanced Hepatocellular Carcinoma / 中国药学杂志

OBJECTIVE: To establish a pharmacodynamics testing and predicting model for molecular targeted therapy of advanced hepatocellular carcinoma (HCC). METHODS: MHCC97-H (a highly aggressive HCC cell line) cells were cultured to establish: ①a subcutaneous tumor model in nude mice, ②an in situ HCC tissue seeding model, ③a hepatic portal vein injection of HCC cell model. Nude mice were intragastric admistrated with 2 mg•kg-1 of sorafenib, apatinib or anlotinib per two days. The antitumor effect of sorafenib, apatinib and anlotinib was examined. RESULTS: Sorafenib, apatinib and anlotinib could significantly inhibit the growth of HCC cells in different models, and the effect of apatinib and anlotinib was better than that of sorafenib. CONCLUSION: By establishing tumor models of HCC in nude mice, this work provides a strategy to examine the potential antitumor activation of agents for advanced HCC.

Effect of deguelin on proliferation of non-small cell lung cancer cell in vitro and in vivo / 中国药理学通报

Aim To investigate the effect of deguelin on the proliferation of non-small cell lung cancer cell line A549 and nude mice. Methods CCK-8 assay was used to detect the inhibition of deguelin on proliferation of SH-SY5Y cells; Hoechst stains and Annex-inV-FITC/PI double stained method were employed to observe the apoptotic morphology and apoptotic rate; flow cytometry was applied to determine the effect of deguelin on cell cycle of A549; tumor xenograft experiment and HE staining were conducted to investigate the effect of deguelin on growth of transplanted tumor in nude mice. Results Deguelin inhibited cell proliferation of A549 dose- A nd time-dependently; Hoechst stains and AnnexinV-FITC/PI double stained further confirmed that deguelin could induce the apoptosis of A549 cells, while deguelin blocked A549 cell cycle in G2/M phase in concentration-dependent manner. The nude mice xenograft model and HE staining experiments showed that deguelin significantly inhibited the growth of xenografts in A549 nude npce (P < 0. 01). Conclusions Deguelin has a significant inhibitory effect on non-small cell lung cancer cell line A549, pointing to a basis for the study of the antitumor activity of deguelin.

Transauricular arterial access for hepatic artery embolization in rabbit VX2 liver tumor

Purpose: The purpose of this study is to evaluate the feasibility of percutaneous transauricular artery access for hepatic artery catheterization using a peripherally inserted central catheter (PICC) device and hepatic artery catheterization through auricular approach. Methods: Ten New Zealand White rabbits were used to establish a VX2 liver tumor model. Hepatic artery angiography and embolization were performed 3 weeks after inoculation. The rabbits were restrained in supine position under anesthesia. Intra-arterial access was accomplished with percutaneous Seldinger technique through the auricular artery using a PICC device. The hepatic artery catheterization was performed with a microcatheter and guide wire. The rate of technical success and procedure time was investigated. Results: Two rabbits failed initial percutaneous transauricular arterial access, with success in a contralateral attempt. Thus, percutaneous transauricular arterial access was achieved in 10 of 12 auricular arteries, with a technical success rate of 83.3%. The time needed to obtain intra-auricular access was 7.2 ± 3.1 min. Hepatic artery catheterization, angiography, and embolization were accomplished through the auricular approach in all 10 rabbits. Conclusion: Arterial access in rabbits can be achieved through the auricular artery. Hepatic artery catheterization, angiography, and embolization can be performed through auricular arterial access

Aquatic Exercise at Thermoneutral Water Temperature Enhances Antitumor Immune Responses

Despite the broad rehabilitative potential of aquatic exercises, the relationship between aquatic exercise and the immune system has not been fully elucidated to date. In particular, there are few specific and delicate immunological approaches to the effect of water temperature on immunity. Thus, we examined the effect of water temperature on immunity during aquatic exercise. The animal tumor model was adopted to examine the impact of aquatic exercise at thermoneutral temperature (TT; 29°C) on immunity compared with aquatic exercise at body temperature (BT; 36°C). Tumor-bearing mice were made to swim in TT water or in BT water for 3 wk and immune cells and their functional activity were analyzed using FACS. Tumor growth was significantly suppressed in mice that exercised in TT than in BT water. The tumor control correlated with the increased number of NK (2-fold), γδT cells (2.5-fold), NKT (2.5-fold), and cytotoxic CD8⁺ T cells (1.6-fold), which play a critical role in anti-tumor immune responses. Furthermore, the functional activity was dramatically improved in the TT group, showing enhanced production of IFNγ in CD8⁺ T cells compared with the BT group. This study demonstrates that aquatic exercise in TT water may improve protective immune responses more effectively than in BT water. Although the effects of water temperature on immune function need further verification in humans, this study suggests that water temperature in human hydrotherapy may be important for improving immune function.

The application and prospects of organoid technique in esophageal cancer research / 解放军医学杂志

Esophageal cancer is a common gastrointestinal tumor. The morbidity and mortality of esophageal cancer in China were 18.85/100,000 and 14.11/100,000, respectively. China is one of the regions with high incidence and high mortality of esophageal cancer in the world. Among them, esophageal squamous cell carcinoma is more common with difficult of early diagnosis. Therefore, there is a great space for research and exploration of early diagnosis and early treatment. Many factors restrict the research on the developing mechanism, drug discovery and screening as well as individualized treatment of esophageal cancer. The lack of an ideal tumor model is one of the important restrictive factors. Organoid is a novel product of 3D culture technology in vitro with the advantages of maintaining the structure, function, genome and drug sensitivity of in situ tissue, and is an excellent tumor research model with such advantages as easy to be operated, relatively low cost, and can be used in conjunction with other advanced technologies. Therefore, esophageal cancer organoid has gradually gained the attention of the academic world, and is expected to be widely used in the field of esophageal cancer research. The present review focuses on the applications and prospects of organoid in the research of esophageal cancer.

Comparison of responsiveness to cancer development and anti-cancer drug in three different C57BL/6N stocks / 한국실험동물학회지

In our efforts to understand the systemic features of tumors, the importance of animal models is increasing due to the recent growth in the development of immunotherapy and targeted therapies. This has resulted in increased attention towards tumor animal models using C57BL/6N, which are mainly used in immunological studies. In this study, the C57BL/6NKorl stock and two other commercial stocks (C57BL/6NA and C57BL/N6B) are evaluated by comparing the occurrence of tumors using the syngeneic model; furthermore, we compare the response to anti-cancer drugs in the syngeneic model by evaluating survival, growth of tumors, proliferation and molecular biology analysis. In the syngeneic model using LLC (Lewis lung carcinoma) cells, the survival of mice and growth of the tumor showed a better response in the C57BL/6NKorl stock, and was dependent on the cell concentration of the dosing tumor, as compared to the other C57BL/6N stocks. However, the Ki-67 staining showed only little difference in cell proliferation within the tumor tissue each mouse stocks. Comparing the sensitivity to anti-cancer drug by examining changes in growth, volume and weight revealed that cisplatin treatment for tumor-bearing C57BL/6NKorl was more dependent on concentration. The Ki-67 staining, however, showed no difference among the C57BL/6N stocks after cisplatin treatment. The expressions of p27 and p53 tumor suppressor proteins, caspase-3 and Bax showed dose-dependent increase after exposure to cisplatin, whereas the expression of Bcl-2 was reduced in a dose-dependent manner. Furthermore, the expressions of MMP-2 and VEGF involved in metastasis, as well as inflammatory genes IL-1β, IL-6 and IL-10, showed dose-dependent decrease in tumor tissue after cisplatin exposure. Differences observed among the C57BL/6N stocks were not significant. Taken together, our studies reveal that C57BL/6NKorl has the potential of being a useful biological resource established in Korea, as it does not differ from the two commercially available C57BL/6N stocks when considering response to tumor generation and sensitivity to anti-cancer drugs using the syngeneic tumor model.

Establishment of a C57BL/6 J mouse model of metastatic melanoma in the lung / 中国实验动物学报

Objective To investigate the influencing factors involved in the establishment of a C57BL/6 J model of metastatic melanoma in the lung,including the way of tumor inoculation,the number of inoculated cells and the time of tumor formation. Methods Mouse melanoma B16F10 cells were cultured in vitro. 1)Eighteen healthy male C57BL/6 J mice were randomly divided into three groups. Mice in each group received 100 μL cell suspension(including 3 ×106 melanoma cells)via intravenous,intraperitoneal and subcutaneous injection,respectively. After two weeks,the mice were killed and dissected,and the tumor growth and metastasis were observed. 2)Eighteen male mice were randomly divided into three groups. Mice in each group were injected with 3 ×106cells,1 ×106cells, and 3 ×105cells through the tail vein,respectively. After two weeks,mice were killed and dissected,and the tumor growth and metastasis were observed. 3)Eighteen male mice were randomly divided into three groups. Mice in each group were injected with 1×106cells though the tail vein. Mice were killed and dissected after one week, two weeks and three weeks, respectively. The growth and metastasis of tumor were observed. Results 1)The success rate of lung metastasis was 100% in the mice with intravenous injection,but not in mice receiving intraperitoneal injection and subcutaneous injection. 2)The size of metastatic melanoma nodules were moderate in mice inoculated by 1 ×106cells. The number of melanoma metastatic foci was too high in the mice inoculated with 3 ×106cells,but too low in the mice inoculated with 3 ×105cells. 3)Significant metastatic melanoma foci were observed in the mice killed and dissected after two weeks with no death. The number of melanoma foci in the lung was too high in the mice killed after three weeks,while was too low in the mice killed at one week after tumor cell inoculation. Conclusions Intravenous injection of 1×106mouse melanoma cells into C57BL/6 J mice and killed after two weeks is an optimal method for establishment of a mouse model of metastatic melanoma in the lung, and is worth of recommendation.

Application of FTS and NIRF conjugated compounds in imaging and therapy of mouse tumor models / 中国实验动物学报

Objective To study the tumor targeting ability and application of farnesylthiosalicylic Acid (FTS) and heptamethine carbocyanine fluorescent dye-mediated near-infrared imagine in living animals, and confirm the inhibitory effect of this compound on growth of tumor cells.Methods Human breast cancer cell line MCF-7, glioma cell line U251 and prostate cancer cell line PC3 were cultured to logarithmic growth phase, and different concentrations of FTS and FTS-IR783 were added, respectively.We observed the inhibitory effect of those two compounds on the growth of tumor cells.Under fluorescence microscopy, specific accumulation of FTS-IR783 in these tumor cells was observed.The tumor cells (1×106) were transplanted subcutaneously into nude mice.These mice were subjected to intraperitoneal injection of FTS-IR783 (10 nmol/mouse) two weeks later.In the in vivo imaging, near infrared fluorescence signal and tumor volume were measured and their correlation was analyzed.Results Compared with FTS, FTS-IR783 significantly inhibited the growth of MCF-7, U251 and PC3 cells in vitro.FTS-IR783 was specifically uptaken by these three kinds of tumor cells, showing strong near infrared fluorescence in cell agglomerates.After subcutaneous injection of FTS-IR783, the correlation between fluorescence intensity and tumor volume was 0.987, 0.998 and 0.971, respectively.Conclusions The compound of FTS conjugated with near infrared fluorescent dye IR-783 can specifically recognize tumor cells, in both in vitro and in vivo imaging.At the same time, the compound can significantly inhibit the growth of tumor cells, and may be expected to become a new potential targeted drug.

Immunotherapy experimental study of γδT cells from human peripheral blood on human liver cell nude mouse xenograft in vivo / 中国免疫学杂志

Objective:To study the immunotherapy effects of different doses of human peripheral blood γδT cells on human hepatoma cells (SMMC-7721) xenograft model.Methods: (1)The nude mouse model of liver cancer was established by inoculated BALB/c mouse subcutaneous with human hepatoma cell line (SMMC-7721).(2)The mononuclear cells in healthy human were extracted from peripheral blood,and specific amplification γδT cells in vitro.(3) The nude mouse model divided into 5 groups by random.The positive control group was 5-Fu,negative control group was normal saline(NS).The treatment group was injected different doses of γδT cells(1×105,5×105 and 25×105)by nude mice tail vein.The positive control group injected 5-Fu by enterocoelia,negative control group injected NS by tail veins.The inhibition effect of different dose γδT cells on tumor was observed,including weight,food intake and growth conditions,etc.and the changes of tumor volume (TV),relative tumor volume (RTV)and relative tumor appreciation rate[T/C(%)] were compared with positive control group and negative control group.Results: Different dose of γδT cells had different degree of inhibition on nude mouse xenograft growth.RTV compared with saline negative control group was statistically significant (P<0.05).Compared with the positive control group of 5-Fu,the TV growth was significantly lower than the 5-Fu,degree of inhibition was similar in RTV each dose group,and all slightly higher than the 5-Fu positive control group.The each dose group of T/C (%)was slightly lower than the relative tumor proliferation rate of the control group of 5-Fu,but had no significant difference.Conclusion: The γδT cells from peripheral blood had significant inhibitory effect on nude mice transplanted liver tumor and it may be used as a new treatment for liver cancer immunotherapy provide experimental data.

Metallothinein 1E Enhances Glioma Invasion through Modulation Matrix Metalloproteinases-2 and 9 in U87MG Mouse Brain Tumor Model

Malignant glioma cells invading surrounding normal brain are inoperable and resistant to radio- and chemotherapy, and eventually lead to tumor regrowth. Identification of genes related to motility is important for understanding the molecular biological behavior of invasive gliomas. According to our previous studies, Metallothionein 1E (MT1E) was identified to enhance migration of human malignant glioma cells. The purpose of this study was to confirm that MT1E could modulate glioma invasion in vivo. Firstly we established 2 cell lines; MTS23, overexpressed by MT1E complementary DNA construct and pV12 as control. The expression of matrix metalloproteinases (MMP)-2, -9 and a disintegrin and metalloproteinase 17 were increased in MTS23 compared with pV12. Furthermore it was confirmed that MT1E could modulate MMPs secretion and translocation of NFkB p50 and B-cell lymphoma-3 through small interfering ribonucleic acid knocked U87MG cells. Then MTS23 and pV12 were injected into intracranial region of 5 week old male nude mouse. After 4 weeks, for brain tissues of these two groups, histological analysis, and immunohistochemical stain of MMP-2, 9 and Nestin were performed. As results, the group injected with MTS23 showed irregular margin and tumor cells infiltrating the surrounding normal brain, while that of pV12 (control) had round and clear margin. And regrowth of tumor cells in MTS23 group was observed in another site apart from tumor cell inoculation. MT1E could enhance tumor proliferation and invasion of malignant glioma through regulation of activation and expression of MMPs.

Influence on radiosensitivity of lung glandular cancer cells when ERCC1 gene silenced by targeted siRNA

Objective To identify the influence on radiosensitivity of lung glandular cancer cells when excisions repair cross-complementing group1 (ERCC1) gene was silenced by targeted siRNA. Methods siRNA which targeting to ERCC1 and control siRNA was designed and synthesized. The human lung glandular cancer SPC-A-1 cells was transfected. A total of 56 nude mice were divided into two groups, and two kinds of SPC-A-1 cells were transplanted to armpit of right forelimb, to establish the nude mice subcutaneous xenotransplanted tumor model of human lung glandular cancer cells. After the tumor was developed, the nude mice were randomly divided into four groups and accepted different doses of X-Ray radiation, then the change of tumor volume, survival time of mice in every group were recorded and the average lifetime was calculated. Twenty-one days later of X-ray experiment, two mice were taken and killed in each group and the tumors organizations were stripped. The cell apoptosis rate and cell cycle distributions were obtained by FCM (flow cytometry). Results The volume of tumor which ERCC1 gene was silenced was less than single irradiation group after X-ray irradiation, and the growth speed was slower and the lifetime of mice was lengthened as well (P < 0.05). The cells apoptosis rate and the rate of G

Study on the influence of curcumin on chemosensitivity of nephroblastoma cells

Objective To study the influence of curcumin on chemosensitivity of nephroblastoma cells. Methods Human nephroblastoma cells line SK-NEP-1 was transplanted to the nude mice subcutaneously to establish the implantation tumor model of human nephroblastoma cells. A total of 30 tumor-bearing mice were divided into three groups of ten randomly. The routine chemotherapy group was given vincristine (0.05 mg/mL·0.2 mL/d) and actinomycin D (15 ng/mL·0.2 mL/d) combined chemotherapy regime. The curcumin chemotherapy group was given the same combined chemotherapy regimens and curcumin (30 mg/kg/d) by intraperitoneal injection. The control group was given normal saline (NS) of the same volume by intraperitoneal injection. Continuous administration would be kept for 4 weeks and 3 days a week. The volumetric changes of every group were recorded. The serum of every group in different time was collected and the VEGF content was detected by ELISA. All mice were cercrificed and the tumor tissues were stripped and weighed after 4 week's treatment. The tumor inhibition rate was calculated. The cell proliferation activity and apoptosis rate were detected by MTT and flow cytometry method. All data were statistically analyzed by SPSS 19.0. Results The tumor volume, serum VEGF content, tumor inhibition rate, cell proliferation activity and apoptosis rate of routine chemotherapy group and curcumin chemotherapy group had significant differences comparing with the control group (P < 0.05) after 4-week's treatment. The cancer growth of curcumin chemotherapy group was obviously decreased and even tended to shrink comparing with routine chemotherapy group (χ

Protective effect of antioxidant on renal damage caused by doxorubicin chemotherapy in mice with hepatic cancer

Objectives To investigate the protective effects and mechanism of antioxidant TBHQ on renal damage caused by doxorubicin chemotherapy in mice with hepatic cancer. Methods Cell H22 of mice with hepatic cancer which was subcultured for three times was subcutaneously transplanted to the groin of right lower limb of 45 SPF Kunming mice to establish the transplanted tumor model. The doxorubicin chemotherapy group and antioxidant intervention group received intraperitoneal injection of ADM (1 mg/kg·0.2 mL/2 d). The model control group received normal saline (NS) of the same volume at the same time. 1% TBHQ was added into the diet of mice of the antioxidant intervention group. Seven weeks later, morning urines and peripheral blood were randomly collected to detect UAlb, UCr, BUN, Scr and UAlb/Cr levels. All mice were beheaded. The renal tissues were made into homogenate, and SOD, T-AOC and MDA content in tissues were detected followed by cell lysis. All data were processed using SPSS19.0. Results The UAlb/Cr, BUN, Scr and MDA of doxorubicin chemotherapy group were significantly higher those of model control group and the activities of SOD, T-AOC in doxorubicin chemotherapy group were lower than those of model control group (P < 0.01). The UAlb/Cr, BUN, Scr and MDA of antioxidant intervention group were lower than those of doxorubicin chemotherapy group and the activities of SOD, T-AOC of antioxidant intervention group were higher than those of doxorubicin chemotherapy group doxorubicin chemotherapy group (P < 0.05). The BUN of model control group was higher than that of blank group, and T-AOC was lower than that of blank group, and difference was statistically significant (P < 0.05). Conclusions Doxorubicin chemotherapy could lead to abnormal antioxidant capacity and renal function of tumor-bearing mice with hepatic cancer. TBHQ antioxidant intervention could effectively improve the antioxidant capacity of renal tissue and reduce the renal damage caused by doxorubicin to some extent.

Influence on radiosensitivity of lung glandular cancer cells when ERCC1 gene silenced by targeted siRNA

OBJECTIVE@#To identify the influence on radiosensitivity of lung glandular cancer cells when excisions repair cross-complementing group1 (ERCC1) gene was silenced by targeted siRNA.@*METHODS@#siRNA which targeting to ERCC1 and control siRNA was designed and synthesized. The human lung glandular cancer SPC-A-1 cells was transfected. A total of 56 nude mice were divided into two groups, and two kinds of SPC-A-1 cells were transplanted to armpit of right forelimb, to establish the nude mice subcutaneous xenotransplanted tumor model of human lung glandular cancer cells. After the tumor was developed, the nude mice were randomly divided into four groups and accepted different doses of X-Ray radiation, then the change of tumor volume, survival time of mice in every group were recorded and the average lifetime was calculated. Twenty-one days later of X-ray experiment, two mice were taken and killed in each group and the tumors organizations were stripped. The cell apoptosis rate and cell cycle distributions were obtained by FCM (flow cytometry).@*RESULTS@#The volume of tumor which ERCC1 gene was silenced was less than single irradiation group after X-ray irradiation, and the growth speed was slower and the lifetime of mice was lengthened as well (P < 0.05). The cells apoptosis rate and the rate of G2/M cells which ERCC1 gene was silenced were higher than the same dose control group and the rate of G1 cells were lower, which indicated that the cells could be stopped at G2/M point, the cell proliferation was inhibited, the cell apoptosis was promoted and the radiation sensitivity was improved after the ERCC1 was silenced.@*CONCLUSIONS@#The radiation sensitivity of lung glandular tumor could be improved after the ERCC1 gene was silenced by siRNA.

Study on the influence of curcumin on chemosensitivity of nephroblastoma cells

OBJECTIVE@#To study the influence of curcumin on chemosensitivity of nephroblastoma cells.@*METHODS@#Human nephroblastoma cells line SK-NEP-1 was transplanted to the nude mice subcutaneously to establish the implantation tumor model of human nephroblastoma cells. A total of 30 tumor-bearing mice were divided into three groups of ten randomly. The routine chemotherapy group was given vincristine (0.05 mg/mL·0.2 mL/d) and actinomycin D (15 ng/mL·0.2 mL/d) combined chemotherapy regime. The curcumin chemotherapy group was given the same combined chemotherapy regimens and curcumin (30 mg/kg/d) by intraperitoneal injection. The control group was given normal saline (NS) of the same volume by intraperitoneal injection. Continuous administration would be kept for 4 weeks and 3 days a week. The volumetric changes of every group were recorded. The serum of every group in different time was collected and the VEGF content was detected by ELISA. All mice were cercrificed and the tumor tissues were stripped and weighed after 4 weeks' treatment. The tumor inhibition rate was calculated. The cell proliferation activity and apoptosis rate were detected by MTT and flow cytometry method. All data were statistically analyzed by SPSS 19.0.@*RESULTS@#The tumor volume, serum VEGF content, tumor inhibition rate, cell proliferation activity and apoptosis rate of routine chemotherapy group and curcumin chemotherapy group had significant differences comparing with the control group (P < 0.05) after 4-week's treatment. The cancer growth of curcumin chemotherapy group was obviously decreased and even tended to shrink comparing with routine chemotherapy group (χ(2) = 15.732, P = 0.007). The cell proliferation activity was significantly reduced and the apoptosis rate was significantly higher, (χ(2) = 9.427, P = 0.012) which showing the effect of chemotherapy was enhanced.@*CONCLUSIONS@#The chemosensitivity of nephroblastoma cells could be improved by curcumin, then the effect of preoperative adjuvant chemotherapy scheme would be enhanced, the growth of nephroblastoma cells would be inhibited and the surgical risk of nephroblastoma would be reduced.

Protective effect of antioxidant on renal damage caused by doxorubicin chemotherapy in mice with hepatic cancer

OBJECTIVES@#To investigate the protective effects and mechanism of antioxidant TBHQ on renal damage caused by doxorubicin chemotherapy in mice with hepatic cancer.@*METHODS@#Cell H22 of mice with hepatic cancer which was subcultured for three times was subcutaneously transplanted to the groin of right lower limb of 45 SPF Kunming mice to establish the transplanted tumor model. The doxorubicin chemotherapy group and antioxidant intervention group received intraperitoneal injection of ADM (1 mg/kg·0.2 mL/2 d). The model control group received normal saline (NS) of the same volume at the same time. 1% TBHQ was added into the diet of mice of the antioxidant intervention group. Seven weeks later, morning urines and peripheral blood were randomly collected to detect UAlb, UCr, BUN, Scr and UAlb/Cr levels. All mice were beheaded. The renal tissues were made into homogenate, and SOD, T-AOC and MDA content in tissues were detected followed by cell lysis. All data were processed using SPSS19.0.@*RESULTS@#The UAlb/Cr, BUN, Scr and MDA of doxorubicin chemotherapy group were significantly higher those of model control group and the activities of SOD, T-AOC in doxorubicin chemotherapy group were lower than those of model control group (P < 0.01). The UAlb/Cr, BUN, Scr and MDA of antioxidant intervention group were lower than those of doxorubicin chemotherapy group and the activities of SOD, T-AOC of antioxidant intervention group were higher than those of doxorubicin chemotherapy group doxorubicin chemotherapy group (P < 0.05). The BUN of model control group was higher than that of blank group, and T-AOC was lower than that of blank group, and difference was statistically significant (P < 0.05).@*CONCLUSIONS@#Doxorubicin chemotherapy could lead to abnormal antioxidant capacity and renal function of tumor-bearing mice with hepatic cancer. TBHQ antioxidant intervention could effectively improve the antioxidant capacity of renal tissue and reduce the renal damage caused by doxorubicin to some extent.

Establishment of a mouse model of early lung adenocarcinoma / 中国实验动物学报

Objective To establish a mouse model of early lung adenocarcinoma to serve the imaging studies of early lung adenocarcinoma .Methods Two-hundred and ten 4-week old SPF female Kunming mice were randomly divided into 5 experimental groups (40 mice per group) and 1 control group (10 mice).The mice of experimental groups were subcutaneously injected with a dose of 0.2 ml 1-methyl-3-nitro-1-nitroso-guanidine ( MNNG) solution in concentration of 2.0 mg/mL weekly for 1, 2, 4, 8, 12 weeks (group A-E), respectively, while the mice of control group was subcutane-ously injected with 0.2 mL saline weekly for 12 weeks.Ten mice were randomly sacrificed from each experimental group at the 60th, 80th, 100th and 120th days after initial injection ( group A60 , group A80 , group A100 , group A120; group B60 ,……and so on) , and the bilateral lungs were dissected .The tumor occurrence rate , number and size of lung tumors were observedandrecorded.Results 1.Thetumoroccurrencerateandnumberoflungtumorswerepositivelyproportionalto the time and dose of MNNG injection .No tumor was found in any group at the 60th day.2.At the 80th day, the tumor oc-currence rate in the groups A, B, C, D and E was 0, 10%, 30%, 40%and 50%, respectively.The number of tumors in each group was 0, 4, 42, 60 and 81, respectively.The number of tumors smaller than 0.5 mm in diameter was 0, 4, 25, 31 and 40, respectively.3.At the 100th day, the tumor occurrence rate was 20%, 40%, 100%, 100%and 100%, re-spectively.The number of tumors was 6, 19, 187, 223 and 301, respectively, and the number of tumors smaller than 0.5 mm in diameter was 5, 16, 132, 124 and 123, respectively.4.At the 120th day, the tumor occurrence rate was 30%, 50%, 100%, 100%and 100%, respectively.The number of tumors was 30, 124, 302, 351 and 362 per group, respec-tively.The number of tumors smaller than 0.5 mm in diameter was 21, 98, 123, 140 and 108, respectively.The induced lung adenocarcinomas were confirmed by pathology .Conclusions MNNG solution 0.2 mL (2.0 mg/mL) subcutaneously injected weekly for 4 weeks can produce 100%occurrence rate of lung adenocarcinoma in mice and most tumors are smaller than 0.5 mm in diameter at 100th day after initial injection .It is a most suitable model for imaging studies of mouse early lung adenocarcinoma .