The Research Of The Cell-killing And Sensitizing Effect Of6-gingerol On HepG-2Cell In Chemotherapy In Different Environment

Background:Hepatocellular carcinoma (HCC) is the most common malignanthepatobiliary disease and is responsible for about1million deaths per year, and its incidence will further increase, to reach a plateau in2015-2020. It is estimated that there are about600,000new cases each year, and the death rate are in the third place in all cancers. Currently the treatment of hepatocellular carcinoma include the following, local treatments and systemic treatments. The local treatments includes surgery、transplantation、 percutaneous ablation and radiofrequency、hepatic arterial chemoembolization、selective internal radiation therapy. The systemic treatments includes chemotherapy, immunological and hormonal treatments、targeted therapy、 molecular therapies、et al. In the clinical treatment, we always choose surgery and combination chemotherapy, but the prognosis was poor. Adriamycin (doxorubicin hydrochloride, DOX, adriamyein, ADM), belongs to the anthracycline anticancer drugs, plays an important role in cancer chemotherapy. But,because of its cardiac toxicity, the effect of its chemotherapy was limited. Besides, because of the side effects of liver cancer chemotherapy was significantly high, the high proportion of multi-drug resistant in tumors and the high cost, a considerable proportion of patients who can not tolerate forced treatment interruptions which caused poor efficacy. Therefore, to seek effective anti-tumor methods of lower toxicity and lower price is one of the hot spot of current research.6-gingerol is the main active ingredient of ginger, modern medical studies confirmed that6-gingerol has a series effects of cardiac, prevention and treatment of cardiovascular diseases, anti-clotting, anti-oxidation, anti-tumor, anti-inflammatory, and anti-ulcer effect of stomach, et al. In recent years, domestic and foreign, did a lot of research in the anti-tumor effect of6-gingerol. Series of in vitro experiments shows that,6-gingerol has a broad-spectrum anti-tumor activity, such as liver cancer, ovarian cancer, breast cancer, colon cancer, leukemia inhibit et al. However, its effectiveness and safety needs to be inspection by clinical trials.Objective:This study was purposed to compared the cell-killing and sensitizing effect of6-gingerol on HepG-2cell in normal mode and hypoxia-hypoglycemia mode in chemotherapy.Methods:The HepG-2cells was given by the physiology laboratory of Shandong University. The cell was cultured in DMEM culture medium containing10%FBS,100U/ml penicillin,100μg/ml streptomycin. Then cultured in a37℃, containing5%carbon dioxide humidified incubator. The HepG-2cells was cultured to logarithmic phase and treated with ADM and6-gingerol in different concentrations. Then the CCK-8assay kit was used to determine the proliferation inhibition of HepG-2cells. Cell apoptosis was detected by combining Flow Cytometry and AnnexinV-FITC PI double staining after treated with different drugs. The expression of bcl-2、bax and birc-5in HepG-2cells was detected by RT-PCR assay. The statistics were analysed by SPSS18.0software.Results:A concentrations of6-gingerol and ADM had a certain degree of growth inhibition on HepG-2cells. In two modes, the inhibition ratio of the6-gingerol and ADM both increased with the increase of the concentration, it showed a dose-dependent manner. Flow Cytometry analysis demonstrated that the apoptosis rate of the experimental group was increased compared to the control group under the two culture conditions,the difference statistically significant. The combination group have the highest number of apoptosis and the number of apoptosis was higher in hypoxia-hypoglycemia group than in normal culture group. Real-time PCR analysis showed that:Compared the control group with the experimental group, the expression of bcl-2、birc-5was decreased and bax have no significant changes under the normal culture conditions. The expression of bcl-2and birc-5was significantly decreased under the hypoxia-hypoglycemia conditions. At the same time, bax expression was increased under the hypoxia-hypoglycemia conditions. Under the hypoxia-hypoglycemia environment, the expression of bcl-2was significantly higher and bax, birc-5’s expression was significantly reduced compared to the normal culture conditions. We can see that, bcl-2/bax was significantly increased in hypoxia-hypoglycemia environment.Conclusion:6-gingerol may decreased the effect of inhibits apoptosis of the Survivin protein by reduced the expression of birc-5. The effect generated the cell-killing and sensitizing effect on HepG-2cell in chemotherapy. The performance of this role is more obvious in the hypoxia-hypoglycemia environment.