| Objective: Primary liver cancer is one of the most common malignant tumors in China now,of which about 90% was hepatocellular carcinoma.The world incidence rates of hepatocellular carcinoma was 43.7%,and it was the second major cause of death from cancer in China.The surgical resection was the preferred treatment for liver cancer,however,the outcomes of patients were poor with high recurrence rate.Most patients with liver cancer has been found with termianl stages which was inoperable.Chemotherapy was an important treatment method for patients with liver cancer to prevent recurrence and metastasis,and to prolong sruvival and improve the quality of life.However,patients with liver cancer showed a strong resistance to chemotherapy and the effects of treatment was reduced.Many clinical studies showed that more than 90% of deathes in patients with cancer were closely related to tumor resistance to chemotherapy each year.Therefore,further study of the molecular mechanisms of drug resistance in liver cancer cells may provide a theoretical basis for the liver cancer drug resistance mechanisms and to find new therapeutic targets to treat liver cancer.Many studies indicated that miRNA may take important function in tumor resistance to chemotherapy.Mi R-223-5p was an important member of the miRNA family involved in the process of multiple drug-resistant tumors,however,there was no information on miR-223-5p in liver cancer resistance to chemotherapy.The purpose of present study was to find the effect of paclitaxel on proliferation rate of human hepatoma cell line BEL-7402 and chemotherapy-resistant hepatocellular carcinoma,also the expression level of miR-223-5p in human hepatoma cell line BEL-7402 and in chemotherapyresistant hepatocellular carcinoma respectively.Analysis of the role of miR-223-5p in the process of drug resistance in liver cancer cells to identify the role of miR-223-5p in chemotherapy-resistant hepatocellular carcinoma and to provide the scientific evidence to reduce hepatocellular resistance to chemotherapy.Method:1 Human hepatoma cell line BEL-7402 were routine cultured and chemotherapy-resistant hepatocellular carcinoma BEL-7402/Taxol were set up;2 The impact of paclitaxelon expression level of human hepatoma cell line BEL-7402 and chemotherapy-resistant hepatocellular carcinoma BEL-7402 was obtained by MTT;3 The expression level of miR-223-5p in human hepatoma cell line BEL-7402 and chemotherapy-resistant hepatocellular carcinoma BEL-7402 detected by qRT-PCR technology.4 The expression level of miR-223-5p in BEL-7402/Taxol cells with miR-223-5p inhibitor was detected by qRT-PCR technology and the impact of paclitaxelon expression level was observed by MTT.Results: 1 The impact of paclitaxelon reducing expression level of human hepatoma cell line BEL-7402 and chemotherapy-resistant hepatocellular carcinoma BEL-7402: The inhibition ratio of two cell lines were both increased with the increasing level of paclitaxelon and incubation time(Ptrend<0.05): the cell inhibition ratio were increased with the increased incubation time with the same level of paclitaxelon,and the cell inhibition ratio of chemotherapyresistant hepatocellular carcinoma were stable with the increased incubation time at low level of paclitaxelon.2 The sensitivity of human hepatoma cell line BEL-7402 and chemotherapyresistant hepatocellular carcinoma BEL-7402/Taxol to paclitaxelon: The IC50 level of paclitaxelon on human hepatoma cell line BEL-7402 and chemotherapy-resistant hepatocellular carcinoma BEL-7402/Taxol were 42.81 ug/ml and 363.50 ug/ml respectively after 24 hours.The IC50 level of paclitaxelon on human hepatoma cell line BEL-7402 and chemotherapy-resistant hepatocellular carcinoma BEL-7402/Taxol were 18.14 ug/ml and 100.14ug/ml respectively after 72 hours.The IC50 level on chemotherapy-resistant hepatocellular carcinoma BEL-7402 was significantly higher(P<0.05).3.1 The expression level of miR-223-5p in human hepatoma cell line BEL-7402 and chemotherapy-resistant hepatocellular carcinoma BEL-7402: The results showed the expression level of miR-223-5p were 5.70±0.33 higher in chemotherapy-resistant hepatocellular carcinoma BEL-7402 than in human hepato cell line BEL-7402.3.2 The expression level of miR-223-5p in human chemotherapy-resistant hepatocellular carcinoma BEL-7402 after miR-223-5p inhibitor were transfected: After miRNA-223-5p-inhibitor were transfected after 24 hours,48 hours and 72 hours respectively,the expression level of miR-223-5p were(86.3±6.45)%,(77.5±5.89)% and(53.5±4.57)% in chemotherapy-resistant group(miR-223-5p inhibitor transfected)compared to control groups(nonmiR-223-5p inhibitor transfected).4 The inhibition ratio in control group and chemotherapy-resistant group: was(28.34.86)% after incubation for 48 hours with 50ug/ml of paclitaxelon.The inhibition ratio in control group and chemotherapy-resistant group was(39.73±2.62)% and(60.41±8.42)% after incubation 48 hours with 100ug/ml paclitaxelon.The inhibition ratio in control group and chemotherapy-resistant group was(55.56±7.63)% and(78.62±6.12)% after incubation 48 hours with 200ug/ml paclitaxelon.The inhibition ratio in chemotherapy-resistant group were all significantly higher(P<0.05)with 50ug/ml,100ug/ml and 200ug/ml paclitaxelon respectively.Conclusion:1 Paclitaxelon has injuring effect on human hepatoma cell.2 The chemotherapy-resistant effect may induced by long term stimulation by paclitaxelon in human hepatoma cell3 The high expression level of miR-223-5p in chemotherapy-resistant hepatocellular carcinoma cell line may be involved in the drug resistant process.4 The sensitivity of human hepatoma cell to paclitaxelon may be improved by reducing expression level of miR-223-5p... |
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