Effect Of Bortezomib Combined With5-Fluorouracil On Human Cervical Cancer Cells (HeLa) And It’s Mechanism

Objectives:Cervical cancer, as the most common gynecologic malignancy, is one of the most serious threats to women’s health and quality of life. There were about200,000women dead of the disease all over the world each year. Traditional treatments of cervical cancer are surgery and radiotherapy mainly, some patients can be cured or alleviated, but some of them after surgery or radiotherapy alone are still easily to recur or get metastasis in short time. Cause they are not sensitive to chemotherapy, the prognosis is poor. Therefore, we all have an urgent need to investigate the more effective way to overcome this diseae. Now, it is generally believed that most anticancer drugs can work by inducing apoptosis of tumor cells. Bortezomib is a kind of proteasome inhibitors, can inhibit the chymotrypsin-like and trypsin-like activity of the proteasome activity in vivo, thereby inhibiting the proliferation of tumor cells.5-Fluorouracil (5-FU) is currently being used as an anticancer drug of cervical cancer. The mechanism of5-FU is known to be due to its metabolic conversion to5-fluorouridine-59-triphosphate with subsequent incorporation intoRNA, and/or the formation of5-fluoro-29-deoxyuridine59-monophosphate, a well-known inhibitor of thymidylate synthetase. The paper focused on the impacts of proteasome inhibitor bortezomib (Bortezomib) which have been used commonly by the blood and tumor oncology combined with chemotherapy drug5-FU on hela cells and analyzed its mechanism superficially.Materials and Methods:Hela cells were cultured in DMEM-H, and were put in37℃saturation humidity,5%CO2incubator. Cells were divided into blank control group, simple Bortezomib group, simple5-FU group and the combination group (Bortezomib in combination with5-FU). MTT assayed inhibition rate of each group. Detected the expression of different groups’Bcl-2, Survivn, IGF-1and HIF-1α mRNA in Hela cells through RT-PCR; PI labeled flow cytometry (FCM) analysed cell cycle changes, and AnnexinV/PI double staining method measured apoptosises of different groups.Results:(1) MTT assay displayed Bortezomib and5-FU can inhibit the proliferation of Hela cells, and the effect was proportional to the concentration and the time. the effect was more significant in combined groups (P<0.05);(2) RT-PCR showed that Bortezomib and5-FU can make the Bcl-2, IGF-1mRNA and Survivn mRNA expressions decreased and was directly proportional to the concentration of drugs, more significant inhibition of these mRNA can be seen in combined groups than alone group (P<0.05); Bortezomib and5-FU’s inhibited effect along on HIF mRNA’s expression was not significant, but the effect of them two was much bigger than the control group (P<0.05).(3) The cell cycle showed that Bortezomib and5-FU may reduce the proportion of cells in S phase,and the effect of the combined group was more significant compared with chemotherapy alone group (P<0.05).(4) The results showed that Bortezomib and5-FU may induce apoptosis in Hela cells, combination of them two is more significant than the control groups (P<0.05).Conclusions:The experimental results showed that, Bortezomib and5-Fu together can significantly increase human cervical cancer Hela cell’s apoptosis and reduce the proliferation of hela cells.