Research Of The Influence And Mechanism About Metformin And Insulin Glargine On The Proliferation Of Endometrial Carcinoma Cell Ishikawa

Objective：Metformin is a widely used antidiabetic drug which has been chose to be thefirst-line drug by many authorities, as ADA, EASD, IDF and CDS. And its safety andefficacy are widely known. The recent advance show that metformin could affecttumor and reduce the risk of cancer. But the mechanism of action is still unknown. Tobe a substitution of traditional insulin, insulin analogue could imitate the model ofphysiological insulin secretion and be widely used in clinical. Lone-acting insulinanalogue, like glargine, shares a homology with IGF-1. In vitro, its effect of mitosis iseight times as much as human insulin. Previous studies have suggested that glarginewould induce cancer. In clinical, the combination of Lone-acting insulin analogue andbiguanides is very common. Whether the combination has a positive function on therate of cancer is a good place to think about, except the superiority in glycemiccontrol. In this study, we discussed the effects of metformin, glargine and thecombination on the proliferation, apoptosis and the express of mTOR in cell-lineIshikawa. We analyzed this effect from the level of cell and gene and observedwhether metformin could stop the proliferation of Ishikawa caused by glargine. Weafford theoretical basis for choosing clinical treatment plan and insulin and approachthe mechanism of tumor suppressor for hypoglycemic drugs.Method:1. Cell culture: culture, expand and passaging Ishikawa conventionally.2. Grouping and intervention: we intervening Ishikawa in logarithmic growthperiod with different concentrations and times of metformin, glargine and thecombination. The control group was intervened with culture fluid without drugs.3. CCK-8detecting cell proliferation.4. Flow Cytometer detecting cell apoptosis.5. Fluorescent quantitation detecting the expression quantity of mTOR.6. Statistical analysis: represent the data with, use SPSS17.0to processdata. The multiple groups were compared with ANOVA. The comparison among groups was performed with LSD-t. P＜0.05means a statistically significantdifference.Result:1. The effect of different concentrations of metformin, glargine and thecombination on the proliferation.1.1Compared with controls, insulin glargine had a higher proliferation. Theproliferation was same among different times(P＞0.05). But the proliferationsbetween different concentrations have statistical significance.(P＜0.05)1.2Different levels of metformin intervening different times. Compare withcontrols, the proliferation was lower in metformin. The drop rate was positivelycorrelated with drug level and Intervention time, with statistical significance(P＜0.05).1.3100IU/L glargine intervened Ishikawa in72h and then putting metformin in.Compared with controls and glargine, the proliferation rate declined dramatically.And the drop rate was positively correlated with drug level, with statisticalsignificance(P＜0.05).2. The effect of different concentrations of metformin, glargine and thecombination on the apoptosis rate.Metformin had a higher apoptosis rate than glargine. The high concentrationgroup had a higher rate. Glargine had on difference in drug levels and had moreeffects on late stage apoptotic rate. Metformin could stimulate Ishikawa to proliferatewith or without Glargine and the rate drops was positively correlated with drug level,with statistical significance(P＜0.05).3. The effect of different level of metformin, glargine and the combination onthe mTOR gene expression.Compare with control, glargine could induce the expression of mTOR gene, withstatistical significance(P＜0.05). In the middle and high concentrations of metformin,the mTOR gene expression was lower than control. The highest one was35.7%.Compare with glargine(100Iu/l), the combine had a significantly reduce, about62.9%-101.6%(P＜0.005). And there was no obvioud difference between thecombine and metformin(P＞0.437). Conclusion:1. Metformin could inhibit Ishikawa cell-line prominently in vitro and stimulateits apoptosis. It could reduce the express of mTOR. The decline was about the druglevel and intervene time.2. In vitro, glargine could stimulate the proliferation of Ishikawa and inhibit itsapoptosis rate and it was positively correlated with glargine level. Glargine couldinduce the up regulated the express of mTOR.3. Metformin could reduce the effect of glargine on proliferation andanti-apoptosis and reduce the express of mTOR. The effect was remarkable when themetformin level was high.