西罗莫司对人子宫内膜癌Ishikawa细胞增殖的影响及其机制

目的研究西罗莫司(SRL)对体外培养的人子宫内膜癌细胞系Ishikawa细胞的增殖及其可能的机制。方法用氯化钴(CoCl2)诱导细胞模拟缺氧微环境,并根据CoCl2浓度不同分为0、25、50、100、150μmol/L组;选择CoCl2浓度为100μmol/L来模拟肿瘤内缺氧微环境,同时分别联合不同浓度(0、10、100、500、1 000 nmol/L)SRL作用细胞48 h。应用CCK-8法测定SRL对各组细胞抑制效应和RT-PCR测定各组mTOR、HIF-1αmRNA的相对表达量。结果随着CoCl2作用浓度的增加,Ishikawa细胞中HIF-1αmRNA表达增强(P<0.05),其中100、150μmol/L两组比较差异无统计学意义(P>0.05);缺氧条件下不同浓度的SRL可抑制Ishikawa细胞,且抑制率随药物浓度的增加而上升(P<0.05);缺氧条件下SRL可使Ishikawa细胞中mTOR、HIF-1αmRNA的表达降低并呈剂量依赖性(P<0.05)。结论缺氧微环境可以促进Ishikawa细胞中的HIF-1αmRNA的表达,HIF-1α可能通过在细胞缺氧调控中起作用而参与了肿瘤的发生发展。在缺氧条件下,SRL可通过抑制mTOR而下调人子宫内膜癌Ishikawa细胞内HIF-1α基因表达。这可能是SRL抗肿瘤的机制之一,并有望成为子宫内膜癌分子治疗的有力途径。

Research of Sirolimus on cell proliferation and its mechanism in human endometrial cancer ishikawa cells

Objective To investigate the effect of Sirolimus on the expression of hypoxia-inducible factor-1alpha(HIF-1α) in human Endometrial Cancer Ishikawa Cells induced by cobalt chloride.Methods Cobalt chloride was used to mimic the cell hypoxic microenvironment,and Ishikawa cells were divided into five groups according to different concent rations of cobalt chloride,i.e.0,25,50,100,and 150 μmol/L groups.The concent ration of 100μmol/L of cobalt chloride was chosen to mimic tumor hypoxic microenvironment,and at the same time,different concent rations of Sirolimus(0,10,100,500,and 1000 nmol/L) were applied to treat the cells for 48h.CCK-8 assay was used to determine the inhibition rate of cell growth.Fluorescence Quantitative PCR(FQ-PCR)technique was used to detect the expression of mTOR,HIF-1αmRNA.Results With increasing of concent ration of cobalt chloride,the expression of HIF-1αmRNA increased(P<0.05),and there was no significant difference between 100 μmol/L group and 150 μmol/L group.The expression of mTOR,HIF-1αmRNA of ishikawa cells induced by cobalt chloride was degressive with the effect of Sirolimus,and became dose-dependent(P<0.05).Conclusion The expression of HIF-1αmRNA of Ishikawa cells can be elevated by hypoxic microenvironment,and HIF-1αmay play an important role in the cell hypoxic regulation to participate in tumorous growth.Sirolimus can inhibit the expression of mTOR,HIF-1αmRNA,which may be one of its antineoplastic mechanisms.