| liver cancer is the most common type of malignant tumor in our country, its incidence is only second to lung cancer, stomach cancer, colorectal cancer, ranking fourth, while its mortality rate ranked second in the world. The poor prognosis of HCC and difficult treatment are because that the disease is insidious onset and early difficult diagnosis, shift has occurred in most patients with locally advanced or already diagnosed. For advanced liver cancer, systemic chemotherapy is often used in clinical palliative therapy, but we have not found a standard chemotherapy drugs and solutions so far.At present, oxaliplatin(oxaliplatin, L-OHP, a third-generation platinum-based chemotherapy drug) is the first-line clinical chemotherapy. Research showed that oxaliplatin can inhibit DNA synthesis, by generating DNA hydration derivative action in the tumor cells, chain and within the chain of crosslink formation, resulting in anti-tumor activity. Because it can inhibit proliferation of tumor cells better, oxaliplatin is not only being used for the treatment of colorectal cancer, etc., but also has a good application in systemic chemotherapy for liver cancer.5-FU(Fluorouracil, 5-FU), uesed in clinical over thirty years, has been widely used in the treatment of liver cancer, stomach cancer, ovarian cancer, choriocarcinoma, malignant mole, colon cancer, colorectal cancer. It is commonly used as anti-metabolic anti-cancer drug in clinical and the first choice for gastrointestinal cancer chemotherapy drug. 5-FU blocks the synthesis of RNA and DNA in the tumor cells by a variety of ways,thus playing a role in inhibiting tumor growth.5-FU converted to 5-fluorouridine(5-FUR) in vivo, the incorporation of synthesis, thus playing a role in the respective phase of cells; FU also can inhibit Exosome complex, thus affecting cell’s viability; 5-FU’s main mode of antitumor ribose is phosphorylated and ribosylated in vivo, converted into 5F-dUMP(5- fluorouracil deoxynucleotide), thus inhibiting thymidylate synthase deoxy and preventing dUMP(deoxy-uridine monophosphate) methyl translating into dTMP(deoxythymidine acid), and dTMP is the necessary raw material in synthesis of DNA replication and can lead to cell cycle arrest and apoptosis [1]. However, clinical application found that 5-FU has many shortcomings,for example, it was easy to produce drug resistance, liver cells is not highly sensitive to it[2], oral absorption of famotidine is not complete, it has larger side-effects, dose of toxic and therapeutic is very close, ect. [3-4] but toxic side-effects and lack of efficacy of liver cancer at high doses and other shortcomings limitits its use in clinical. So exploring new chemotherapy regimens to improve the sensitivity of 5-FU on hepatoma cells has become a very important and urgent issue.The new class of anticancer anthraquinone epirubicin was known as epirubicin, compared to doxorubicin, its advantage is more efficient and has a broad spectrum, its part of the 4-amino sugar hydroxyl groups in the original cis style transfers into the present trans.It has lower toxicity than doxorubicin and daunorubicin. At present,it is the first choice among same drugs and has been widely used[4]. Its main mechanism of function is: by inserting between adjacent base pairs, and the DNA double helix forming a reversible binding, interfering with the process of transcription, inhibiting the synthesis of RNA and DNA, in order to achieve the goal of cancer therapy[5]. Zhu Xiaodong et al[6]made a evaluate to the application of epirubicin combined with 5-FU + OXA program for advanced gastric cancer. The results showed that, compared with other chemotherapy regimens, it has a good tolerability and high efficiency, and low toxicity.Objective:By use anthraquinone epirubicin combined with 5-FU and OXA,we increased tumor cell apoptosis rate, whether the treatment of liver cancer can achieve synergistic or additive effect,the joint study of mechanism of three kinds of drugs and scheme optimum requires clinical trials system.Through exoteric experiment,we study on the synergy mechanism of combination of three drugs killing tumor cells,and explore new ideas for the treatment of liver cancer,and provide a theoretical basis for clinical treatment.Methods:1. Cell Source: human hepatoma HepG2 cells were purchased from Shanghai Mei Xuan Biotechnology Co., Ltd.2. Experimental groups: the establishment of epirubicin combined with 5-FU + OXA group, 5-FU + OXA group, epirubicin group, the control group3. MTT to detect the difference activity of liver cancer cell proliferation in treatment groups4. Real-time PCR was used to detect the level of Bcl-2mRNA and BaxmRNA5. Western blot was used to detect the expression of Bcl-2and Bax6. Statistical analysis: SPSS17.0 was used, all data were respected by mean ± standard deviation( x ± s). Multiple comparisons were conducted by ANOVA analysis, Comparisons between two groups were conducted by SNK, P <0.05 suggests statistical meaning.The result:1. MTT was used to detect the effects in different treatment groups on the proliferation of HepG2 cells. Firstly, we screened IC50 of each drug on 48h: epirubicin, 5-FU and OXA IC50 of 3.44 umol /L,1.76mmol/L,6.89mmol/L. The results of MTT showed, four groups with drug intervention HepG2 cells after 48 h, the inhibition rate was 29.80%,(epirubicin group), 22.19%(5-FU + OXA group), 14.86%(epirubicin Star + 5-FU + OXA group), the differences had statistical significance(F = 120.198, P <0.05). Compared with the control group,Epirubicin + 5-FU + OXA group, 5-FU + OXA group, epirubicin comparison group, OD values were lower, each treatment group inhibited the proliferation of HepG2 cells, The differences had statistical significance(P <0.05); In three treatment groups, the OD values in the group of doxorubicin + 5-FU + OXA were the lowest, which suggest it increased the proliferation rate of HepG2 cells effectively(The comparison among the four groups, P <0.05, the differences have statistical significance).2. RT-PCR was used to detect the expression of Bcl-2mRNA and BaxmRNA in Hep G2 cells:compared with control group,epirubicin + 5-FU + OXA group, between 5-FU + OXA group, epirubicin compare doxorubicin group,the expression of Bcl-2mRNA were lower(F = 27.392, P <0.05, the differences had statistical significance), In three treatment groups, the expression of Bcl-2mRNA and BaxmRNA were the lowest in epirubicin Star group(P <0.05, the differences had statistical significance).3. Western Blot was used to detect the expression of Bcl-2mRNA and BaxmRNA in HepG2 cells: compared with the control group, expression of Bcl-2 protein were reduced and Bax were increased in treatment groups,epirubicin +5-FU + OXA group was the most obvious, which suggest that with the combined effects of the drug, the expression of Bcl-2 protein was gradually increased(F = 816.418, P <0.05, the differences had statistical significance), the expression of Bax protein was gradually decreased(F = 62.916, P <0.05, the differences had statistical significance).In conclusion:HepG2 human hepatoma cells in the intervetionof epirubicin combined with 5-FU and OXA, which accelerated apoptosis of tumor cells and d significantly decreased growth rate, the mechanism of combined effects may through inhibit the expression of Bcl-2, thus promoting the expression of Bax. |
PDF Full Text Request