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Objective:To explore the correlation of the dose of capecitabine with the efficacy and cardiotoxicity in patient-derived tumor xenograft (PDX) model of mice with colorectal cancer.Methods:The fresh cancer tissues of 1 colorectal cancer patient were transplanted into the bilateral axillary subcutaneous of immunodeficient NOG mice to establish PDX model and passage stably. And then the morphology of tumor cells in primary generation and the second-generation tumor tissues was observed by using HE staining. The expression of tumor markers was detected by using immunohistochemistry method, and the model was evaluated. Mice were intragastrically infused with 200, 300 and 400 mg/kg capecitabine once a day, which were treated as low, middle and high dose groups respectively, 5 rats in each group; in the control group, 0.9% NaCl solution was perfused into the stomach; 14 d in total, use stop for 7 d, consecutively administered in this way. The body weight was measured every day and the tumor volume was measured every 3 days. After 100 days of observation, the mice were killed, and the tumor tissue was taken to measure the tumor weight and then the tumor volume, tumor volume inhibition rate and tumor inhibition rate were calculated. The morphology of tumor tissues was observed by using HE staining. The protein levels of anti-tumor effect indexes like rasP21, cyclooxygenase 2 (COX2), prostaglandin E2 (PGE2), cardiac troponin Ⅰ (cTn-Ⅰ) and brain natriuretic peptide (BNP) in serum of mice were detected by using enzyme linked immunosorbent assay (ELISA).Results:PDX model of mice with colorectal cancer was successfully constructed, and the histological characteristics of the primary tumor in the model were well preserved. During administration, 1 mouse died in the capecitabine high dose group; a slow down in tumor volume growth could be found with the increased dose of capecitabine. There was no statistically significant difference in body weight among 4 groups until all mice were killed ( P > 0.05). The tumor volume and tumor weight in the low, middle and high dose groups were lower than those in the control group (all P < 0.05), and the tumor volume and tumor weight showed an obvious decrease with the increase in dose. The tumor volume inhibition rates of low, middle and high dose groups were 42.61%, 67.61% and 77.27%, respectively, and the tumor inhibition rates were 35.53%, 67.77% and 75.09%, respectively. The serum anti-tumor effect indexes rasP21, COX2 and PGE2 in the middle and high dose groups were decreased compared with those in the control group (all P < 0.05), while cTn-Ⅰ and BNP levels were increased compared with those in the control group (all P < 0.05). Conclusions:The established PDX model of mice with colorectal cancer can better retain the histological characteristics of the original tumor. After treatment of middle and high dose of capecitabine, the tumor inhibition effect is obvious, but the risk of myocardial damage should be noticed.
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Objective:To investigate the inhibitory effect of brazilin on bladder cancer cells and its mechanism.Methods:Chemically synthesized brazilin was synthesized by chemical synthesis. Methyl thiazolyl tetrazolium (MTT) method was used to detect the inhibitory effect of synthetic brazilin on bladder cancer cells T24 and BIU87. Proteomic technique was used to detect the effect of brazilin on the level of protein in both cells. Real-time quantitative polymerase chain reaction (qRT-PCR) and Western blot methods were used to verify the effects of brazilin on the expression of protein regulator of cytokinesis 1 (PRC1) of both cells at gene and protein level.Results:MTT method showed that brazilin significantly inhibited the proliferation of bladder cancer cells T24 and BIU87, and its half inhibitory concentration ( IC50) of T24 cell and BIU87 cell was 9.9 μg/ml and 5.1 μg/ml,respectively. Proteomic results showed that brazilin could regulate the protein expression of PRC1 in both cells, which was verified by qRT-PCR and Western blot. Conclusion:Brazilin suppresses bladder cancer cell growth possibly by downregulating PRC1.
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Objective:To investigate the inhibiting effect of Sufuning Lotion (SFN) on bladder carcinoma T24 cells.Methods:Trypan blue exclusion test was performed to observe the killing effect of 2 mg/ml SFN at different time points (20, 40, 60, 80, 100 min) on human bladder carcinoma T24 cells; the inhibiting effect of SFN with different concentrations (8.0, 12.0, 18.0, 27.0, 40.5 μg/ml) for 48 h on proliferation of T24 cells was assessed by using methyl thiazolyl tetrazolium (MTT) assay. The half inhibitory concentration ( IC50) was identified. T24 cells were treated with IC50 SFN for 24, 48, 72 h, and then the change of proliferation inhibition rate of T24 cells was detected. The nude mice subcutaneous model (30 mice) and intraperitoneal tumor xenograft model (30 mice) were prepared according to T24 cells inoculated method. After inoculation for 24 h, both animal models were divided into 5 groups with 6 animals in each group based on the random number method, including the control group (0.9% NaCl solution), the SFN 200 mg/kg group, the SFN 300 mg/kg group, the SFN 400 mg/kg group and the mitomycin group, and then the control group and three SFN groups were intraperitoneally injected for 6 d, while the mitomycin 1 mg/kg group was injected with 1 mg/kg mitomycin every 5 d for once, 2 times in total. The transplantable tumor volume of subcutaneous tumor xenograft model was measured per week and the mice were sacrificed after 4 weeks. Tumor tissues were taken out to measure the tumor weight and tumor growth inhibition ratio was also evaluated. The survival time of nude mice in intraperitoneal tumor xenograft model was recorded so as to calculate the life extension rate. Results:Trypan blue exclusion test showed that after the function of 2 mg/ml SFN for 20, 40, 60, 80, 100 min, the cell death rate was (17.83±1.56)%, (48.95±1.34)%, (67.46±1.44)%, (75.48±2.12)%, (89.41±1.35)%, respectively, and the difference was statistically significant ( F = 1 213.264, P < 0.01). MTT assay showed that SFN inhibited the proliferation of T24 cells in a concentration-dependent and time-dependent manner, and the IC50 of cell proliferation at 48 h was (14.36±0.35) μg/ml. After the function of 14.36 μg/ml SFN for 24, 48, 72 h, the proliferation inhibitory rate of T24 cells was (39.5±0.9)%, (50.6±0.7)%, (71.5±1.0)%, respectively, and differences was statistically significant ( F = 1 044.206, P < 0.01). After the nude mice was inoculated with T24 cells for 4 weeks, the tumor volume and tumor weight in the SFN 200 mg/kg group, the SFN 300 mg/kg group, the SFN 400 mg/kg group and the mitomycin group were lower than those in the control group [the tumor volume: (0.925±0.136) cm 3, (0.833±0.171) cm 3, (0.652±0.117) cm 3, (0.482± 0.120) cm 3 vs. (1.231±0.210) cm 3, respectively; the tumor weight: (1.56±0.20) g, (1.42±0.21) g, (1.19±0.22) g, (0.97±0.16) g vs. (1.98±0.30) g], and differences were statistically significant ( F = 20.153, P < 0.01; F = 17.325, P < 0.01); there were no significant differences in the tumor volume and weight between the SFN 400 mg/kg group and the mitomycin group ( t = 1.898, P = 0.069; t = 1.739, P = 0.094), the inhibition rate of subcutaneous tumor xenograft model was 20.94%, 28.28%, 39.66%, 51.14%, respectively in the SFN 200 mg/kg group, the SFN 300 mg/kg group, the SFN 400 mg/kg group and the mitomycin group. The survival time of intraperitoneal nude mice in the SFN groups and the mitomycin group was prolonged compared with that in the control group [(32.7±3.2) d, (34.0±4.5) d, (34.3±2.3) d, (35.3±2.0) d vs. (21.7±4.8) d], and there was a statistically significant difference ( F = 15.179, P < 0.01), the life extension ratio was 50.76%, 56.90%, 58.42%, 63.04%, respectively. Conclusion:SFN can inhibit the proliferation of T24 cells, and it has an anti-tumor effect on the T24-bearing nude mice.
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Objective:To establish a patient-derived xenotransplantation (PDX) animal model of gastric cancer, and observe the anti-cancer effect of chemotherapeutic drugs on this model.Methods:Human gastric cancer tissues were inoculated into the subcutaneous tissues of both axillaries of NOG mice and were subcultured for 3 generations. The tumor tissues of the third-generation NOG mice were selected and inoculated into the subcutaneous tissues of left axillary of 21 severe combined immunodeficiency-non-obese diabetes mellitus (SCID-NOD) mice to establish PDX mouse model of gastric cancer. The inoculated mice were divided into control group (mice received only 0.9% sodium chloride injection), oxaliplatin group, cisplatin group, paclitaxel group, fluorouracil group, tegafur, gimeracil and oteracil porassium capsules group and capecitabine group, with 3 mice in each group, and the corresponding drugs were given. The mice survival status, tumor volume and tumor weight at different times were recorded. Mice were sacrificed on the 61st day of administration, and the tumor inhibition effects of 6 kinds of chemotherapy drugs on the PDX model of gastric cancer were evaluated.Results:After being subcultured for 3 generations, the stability of tumor transmission in PDX animal model of gastric cancer was improved, and the homogeneity of tumor growth was good at the initial stage. At the early stage of administration, the model was more sensitive to oxaliplatin, fluorouracil and capecitabine, and the tumor growth inhibition (TGI) values on the 31st day were 63.37%, 52.11% and 78.48%, at the end of administration, the model had the best sensitivity to capecitabine with a TGI value of 59.22% and a tumor inhibition rate of 58.65% on the 61st day. The TGI curve after administration showed that paclitaxel had no obvious anti-tumor effect, cisplatin had the worst anti-tumor effect, and the model had poor tolerance to tegafur, gimeracil and oteracil porassium capsules.Conclusion:The PDX animal model of gastric cancer is successfully established, and capecitabine has the best tumor suppressive effect on this model.
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Objective To observe the sensitization of lidocaine on subcutaneous hepatoma H22-bearing mice and abdominal cavity H22 tumor-bearing mice treated by mitomycin. Methods According to the random number table method, the mice were divided into subcutaneous tumor-bearing group and abdominal cavity tumor-bearing group, with 15 mice in each group. The mice in the two groups were further divided into three subgroups: model group, mitomycin group, mitomycin+lidocaine group, with 5 mice in each subgroup. The day before the intraperitoneal injection, the density of H22 cells obtained from peritoneal culture of one mouse was adjusted to 5 ×106/ml. Subcutaneous tumor-bearing group mice were injected H22 cells into the right armpit, and abdominal cavity tumor-bearing group mice were injected H22 cells into the abdominal cavity, 0.2 ml per mouse. Intraperitoneal injection was given after inoculation for 24 h (the experiment day 1), followed by intraperitoneal injection on day 5 and 9. Univariate ANOVA analysis and t test were used to analyze the solid tumor weight and tumor inhibition rate on the 11th day of subcutaneous tumor-bearing mice, and the survival time and life extension rate within 60 days of abdominal cavity tumor-bearing mice. Results The solid tumor weight of subcutaneous tumor-bearing mice model group, mitomycin group and mitomycin + lidocaine group were (3.77 ±1.02) g, (1.67 ±0.28) g, (0.74 ±0.19) g, respectively, and the differences in the three groups were statistically different (F = 31.753, P < 0.01); compared with the subcutaneous model group, the subcutaneous solid tumor weights of mitomycin group and mitomycin +lidocaine group were decreased and the differences were both statistically different (t=2.10, P<0.01; t=3.04, P<0.01); the subcutaneous solid tumor weight of mitomycin+lidocaine group was lower than that of mitomycin group (t= 0.93, P= 0.034). The tumor inhibition rate of mitomycin group and mitomycin +lidocaine group reached 55.70％ and 80.37％ respectively. The survival time of abdominal cavity tumor-bearing mice in model group, mitomycin group and mitomycin + lidocaine group was (16.80±0.84) d, (28.80± 6.30) d, (40.40±12.86) d, respectively, and the differences in the three groups were statistically different (F=10.155, P=0.003); compared with the abdominal cavity tumor-bearing mice model group, the survival time of mice in mitomycin group and mitomycin + lidocaine group was prolonged (t= 12.00, P= 0.041; t= 23.60, P= 0.001), and it was found that survival time in mitomycin + lidocaine group was longer than that in mitomycin group (t=11.60, P=0.047). The life extension rate of mitomycin group and mitomycin+lidocaine group reached 71.43％ and 140.48％ respectively. Conclusion Lidocaine can increase the sensitization of mitomycin on hepatoma H22-bearing mice.
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Objective To investigate the efficacy and safety of intravesical instillation with Sufuning (SFN) lotion for prevention of postoperative recurrence of bladder cancer. Methods A total of 240 bladder cancer patients who were diagnosed as bladder cancer and accepted trans-urethral resection of bladder tumor from January 2010 to June 2016 in Shanxi Provincial Cancer Hospital were randomly divided into the experimental group (120 cases) and the control group (120 cases) according to the envelope method. The patients in the experimental group were treated with SFN lotion for immediate intravesical instillation(250 mg for once), and the patients in the control group were treated with pirarubicin (THP) for immediate intravesical instillation (30 mg for once). The patients of two groups were treated with intravesical chemotherapy once a week for 8 times, and the chemotherapy was performed once a month for 1 year. The recurrence rate, progression-free survival (PFS) rate, overall survival (OS) rate and recent side effects were compared between the two groups. Results The patients were followed up for 6 to 60 months. The median follow-up time was 36.5 months.In the experimental group,6 patients were lost and 8 patients were lost in the control group.The experimental group, the total recurrence rate was 26.3 % (30/114). The control group, the overall recurrence rate was 25.0 % (28/112) (χ2= 0.142, P = 0.781). Five years of PFS rate in the experimental group and the control group was 73.7 % (84/114) and 75.0 % (84/112) respectively, and there was no significant difference between the two groups (χ2= 2.011, P= 0.615). Five years of OS rate in the experimental group and the control group was 95.6 % and 92.9 % respectively, and there was no significant difference between the two groups (χ 2= 1.611, P= 0.425). The major side effects included chemical cystitis and hematuria. The incidence of chemical cystitis and hematuria in the experimental group was significantly lower than that in the control group(χ2=5.991,P=0.018;χ2=4.925,P=0.036).There was a statistically significant difference of the hematological side effects (blood routine changes) between the two groups (χ 2= 4.891, P= 0.032). Conclusion It is safe and effective for intravesical instillation of SFN lotion to prevent the recurrence of bladder cancer.
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Objective To establish a stable animal model of transplanted human colorectal cancer.Methods BALB/c nude mice were randomly divided into orthotopic colorectal model group and subcutaneous inoculation model group by random number table,and separately inoculated with 0.1 ml human colon cancer cell HCT116 under the density of 2×107/ml into the orthotopic colorectal (with the self-made inoculator) and right forelimb pit subcutaneously.The mice were observed for 60 days to compare the tumor formation and tumor growth in the two groups.Results The tumor formation rate of all 18 animals was 100 ％ (18/18) in the orthotopic colorectal group.The average tumor weight was (2.78±1.86) g and the average survival time was (45.00±11.99) d.The tumor formation rate was 27.78 ％ (5/18) in the subcutaneous inoculation group.The average tumor weight was (1.74±0.82) g,and the average survival time was (60.00±0.00) d.Conclusion 0.1 ml (2×107/ml) human colon cancer cell suspension HCT116 inoculated into BALB/c nude mice orthotopic colorectal with self-made inoculator could establish human colorectal cancer animal model successfully.
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Objective To establish a method for preparing orthotropic transplantable hepatocellular carcinoma in mice. Methods According to the liver detailed anatomical structure of the mouse, 50 μl mouse ascites containing 1 ×106 and 5 ×105 mouse hematoma H22 cells was input to liver in 12 Kunming mice through percutaneous intraperitoneal injection by syringe, respectively, to establish orthotopic transplantable hepatocellular carcinoma model. The growth status of mice was observed, and the pathological changes of liver and tumor metastasis tissues were detected. The tumor formation and metastasis were analyzed. Results The tumor formation rate was 100% (12/12) by direct injection of mouse hematoma H22 cells in 2 groups. The inoculated mice started to appear ascites at the 6th day, and all mice produced ascites at the 10th day. The survival time was (16.17 ±3.07) d and (18.08 ±3.34) d in 1 ×106 group and 5 ×105 group, respectively. Some mice emerged tumor metastasis in kidney, intestine, spleen and mesenteric lymph nodes. Conclusion The method of direct injection could establish orthotropic transplantable hepatocellular carcinoma model in mice, which can be used for antitumor drug research.
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Objective To observe the effects of brazilin on proliferation and apoptosis in T24 cells.Methods Trypan blue exclusion test was performed to detect the inhibition of brazilin on the growth of T24 cell lines in vitro cultured within different time.After exposure to different concentrations of brazilin,homogeneous bioluminescence assay was used to detect the inhibitory action of brazilin,Caspase-3 and Caspase-9 activity on T24 cells.Cellular apoptosis was measured by flow cytometry (FCM) and observed by laser scanning confocal microscope.Results Brazilin could significantly inhibit the proliferation of T24 cells after 8 hours,the inhibitory rates of the brazilin at concentration of 25,50,100,200 μg/ml against T24 cells respectively were 43.19 ％,60.73 ％,86.38 ％ and 93.89 ％ (P ＜ 0.05).After exposured to 50 μg/ml of brazilin,the inhibition ration to T24 cells increased with time prolonging (52.72 ％ in 4 h,60.73 ％ in 8 h,91.77 ％ in 24 h,96.41％ in 48 h) (P ＜ 0.05).The activity of Caspase-3 and Caspase-9 increased slightly when brazilin was at 25 μg/ml,but there was no statistical differences compared with that in the control group (P ＞ 0.05).When cells were treated with an increase of the concentration of brazilin from range of 7.5-60 μg/ml for 16 hours,the apoptosis ratio in turn showed a upward trend of 0.15 ％,1.35 ％,2.91％,34.76 ％.It could be seen by laser scanning confocal microscope that the apoptosis occurred in the cells.Conclusion Brazilin can effectively inhibit the proliferation of T24 cells and induce apoptosis in a dose and time dependent manner.