大黄衍生物B对白介素-6诱导宫颈癌Hela生物学行为的影响

Effect of rhubarb derivative B on biological behavior of cervical cancer Hela induced by IL-6

目的探讨大黄衍生物B通过AMPK/mTOR通路对白介素-6(IL-6)诱导的子宫癌细胞凋亡、迁移及侵袭的影响。方法将宫颈癌Hela细胞分为对照组、IL-6组、高剂量组、中剂量组和低剂量组。IL-6组添加25 ng/ml IL-6,低剂量组添加25 ng/ml IL-6+40μmol/L大黄衍生物B,中剂量组添加25 ng/ml IL-6+80μmol/L大黄衍生物B,高剂量组添加25 ng/ml IL-6+120μmol/L大黄衍生物B。MTT试剂检测宫颈癌细胞增殖,流式细胞仪检测宫颈癌细胞凋亡,细胞划痕测定宫颈癌细胞迁移能力,Transwell小室测定宫颈癌细胞侵袭能力,RT-PCR检测各组细胞中AMPK、mTOR、TSC1和TSC2 mRNA表达,Western blot检测各组细胞AMPK、mTOR、p-AMPK、p-mTOR、TSC1和TSC2表达。结果各剂量组宫颈癌细胞增殖抑制率和凋亡率明显低于IL-6组(P<0.01)。各剂量组宫颈癌细胞细胞划痕愈合率和穿膜细胞数明显低于IL-6组(P<0.01)。各剂量组宫颈癌细胞AMPK、TSC1和TSC2 mRNA表达明显高于IL-6组(P<0.01),mTOR mRNA表达明显低于IL-6组(P<0.01)。各剂量组宫颈癌细胞AMPK、p-AMPK、TSC1和TSC2蛋白表达明显高于IL-6组(P<0.01),mTOR和p-mTOR蛋白表达明显低于IL-6组(P<0.01)。结论大黄衍生物B通过上调AMPK和下调mTOR通路能够明显抑制白介素-6诱导宫颈癌Hela增殖、迁移及侵袭。

Objective To investigate the effect of rhubarb derivative B on apoptosis,migration and invasion of cervical cancer cells induced by interleukin-6(IL-6)through AMPK/mTOR pathway.Methods Hela cells of cervical cancer were divided into control group,induction group,high dose group,middle dose group and low dose group.IL-6 group added 25 ng/ml IL-6,low dose group added 25 ng/ml IL-6+40μmol/L rhubarb derivative B,middle dose medicine group added 25 ng/ml IL-6+80μmol/L rhubarb derivative B,high dose group added 25 ng/ml IL-6+120μmol/L rhubarb derivative B.MTT reagent was used to detect the proliferation of cervical cancer cells.Flow cytometry was used to detect apoptosis of cervical cancer cells.The migration ability of cervical cancer cells was measured by cell scratch.The invasive ability of cervical cancer cells was measured by Transwell chamber.RT-PCR was used to detect the expression of AMPK,mTOR,TSC1 and TSC2 in each group.Western blot was used to detect the expression of AMPK,mTOR,p-AMPK,p-mTOR,TSC1 and TSC2 in each group.Result The proliferation inhibition rate and apoptosis rate of cervical cancer cells in each dose group were significantly lower than those in group IL-6(P<0.01).The wound healing rate and number of penetrating cells in cervical cancer cells in each dose group were significantly lower than those in group IL-6(P<0.01).The expressions of AMPK,TSC1 and TSC2 in cervical cancer cells of each dose group were significantly higher than those of IL-6 group(P<0.01),and the expression of mTOR was significantly lower than that of IL-6 group(P<0.01).The expressions of AMPK,p-AMPK,TSC1 and TSC2 in cervical cancer cells of each dose group were significantly higher than those of IL-6 group(P<0.01),while the expressions of mTOR and p-mTOR were significantly lower than those of IL-6 group(P<0.01).Conclusion Rhubarb derivative B can significantly inhibit the proliferation,migration and invasion of Hela induced by IL-6 by up-regulating AMPK and down-regulating mTOR pathway.