MTA1调控HIF-1α/VEGF通路作用于胰腺癌细胞增殖和迁移能力的研究

Effect of MTA1 on the proliferation and migration of pancreatic cancer cells by regulating HIF-1α/VEGF pathway

目的研究肿瘤转移相关蛋白1(MTA1)对胰腺癌细胞增殖、迁移和缺氧诱导因子(HIF-1α)/血管内皮生长因子(VEGF)信号通路的影响。方法采用Western blot检测MTA1在人胰腺癌组织、癌旁组织组织中的表达。采用酶联免疫吸附(ELISA)检测胰腺癌肿瘤细胞系PDAC-1培养上清液中MTA1浓度。然后加入不同浓度(5.0 ng/ml、10.0 ng/ml、15.0 ng/ml、20.0 ng/ml)的外源性MTA1处理胰腺癌肿瘤细胞系PDAC-1 48 h,MTT法检测细胞增殖。采用MTA1(20.0 ng/ml)和YC-1(50μmol/L)分别单独或联合作用于胰腺癌肿瘤细胞系PDAC-1 48 h,MTT法检测细胞增殖;采用划痕法检测细胞迁移能力;24 h时Western blot检测HIF-1α/VEGF信号通路中的关键蛋白HIF-1α和VEGF蛋白的水平。结果 MTA1在胰腺癌组织中的表达显著高于癌旁组织(P <0.05)。胰腺癌肿瘤细胞系PDAC-1自然生长条件下分泌的MTA1水平随着时间的增加在不断增大,48h后浓度为(0.041±0.003) ng/ml与后续研究中加入的外源MTA1浓度相比,可忽略不计。MTA1浓度在5.0~20.0 ng/ml时,从作用第12小时开始能够显著促进胰腺癌肿瘤细胞系PDAC-1的增殖(P <0.05)具有时间和剂量依赖效应。YC-1能够抑制胰腺癌肿瘤细胞系PDAC-1的增殖和迁移能力。MTA1能够促进HIF-1α和VEGF蛋白的表达,而YC-1能够降低MTA1对HIF-1α和VEGF蛋白表达的促进作用。结论 MTA1可以促进胰腺癌肿瘤细胞系PDAC-1的增殖和迁移,且具有时间和浓度依赖效应,其作用机制可能与HIF-1α/VEGF信号通路相关。

Objective To investigate the effect of tumor metastasis related protein 1(MTA1) on the proliferation,migration and HIF-1α/VEGF signaling of pancreatic cancer cells.Methods the expression of MTA1 in human pancreatic cancer tissues and adjacent tissues was detected by Western blot,and the concentration of MTA1 in the supernatant of PDAC-1 culture of pancreatic cancer cell line was detected by ELISA.Then,the pancreatic cancer cell line PD AC-1 was treated with different concentrations of exogenous MTA1 for 48 h,and cell proliferation was detected by MTT assay.MTA1(20.0 ng/ml) and YC-1(50μmol/L) were applied to pancreatic cancer cell line PDAC-1 for 48 h,respectively,and MTT assay was used to detect cell proliferation.Cell migration ability was detected by scratch method.At 24 h,Western blot was used to detect the levels of HIF-α and VEGF proteins,key proteins in the HIF-1α/VEGF signaling pathway.Results The expression of MTA1 in pancreatic cancer tissue was significantly higher than that of tongue tissue adjacent to carcinoma(P <0.05),pancreatic cancer cell lines under the condition of natural secretion level of MTA1,the results showed that MTA1 concentration with the increase of time,in the growing,after 48 h concentration(0.041±0.003) ng/ml,compared with the concentration of exogenous MTA1 follow-up study to join,negligible.When MTA1 concentration was 5.0-20.0 ng/ml,it significantly promoted the proliferation of PDAC-1 in pancreatic cancer tumor cell line from the12 th hour(P <0.05),with time-dependent and dose-dependent effects.YC-1 inhibited the proliferation and migration of pancreatic cancer cell line PDAC-1.MTA1 can promote HIF-1α and VEGF protein expression,while YC-1 can reduce the effect of MTA1 on HIF-1α and VEGF protein expression.Conclusion MTA1 promotes the proliferation and migration of pancreatic cancer cell line PDAC-1 with time-dependent and concentration-dependent effects,and its mechanism may be related to HIF-1α/VEGF signaling pathway.