上调PHLPP1表达对食管癌细胞增殖、黏附、侵袭和迁移能力的影响

PHLPP1 inhibits proliferation,adhesion,invasion and migration of esophageal cancer Eca109 cells

目的:研究上调PHLPP1表达对食管癌细胞增殖、黏附、侵袭和迁移能力的影响。方法:用qRT-PCR和Western blot法检测食管癌细胞Eca109、EC9706、KYSE510和正常食管上皮细胞HEEC中PHLPP1表达水平。Eca109细胞感染过表达PHLPP1的重组慢病毒(Lv-PHLPP1组),以感染阴性对照重组慢病毒的细胞为Lv-NC组,未行感染的细胞为对照组,qRT-PCR和Western blot法检测3组细胞中PHLPP1的表达,MTT方法测定细胞增殖和黏附能力,Transwell小室检测细胞侵袭和迁移能力,Western blot法检测细胞中增殖细胞核抗原(PCNA)、神经性钙黏附素(N-cadherin)、基质金属蛋白酶1(MMP1)和9(MMP9)、上皮性钙黏附素(E-cadherin)蛋白的表达水平。结果:Eca109、EC9706、KYSE510中PHLPP1表达水平均低于HEEC细胞,Eca109细胞中PHLPP1表达水平最低(P <0. 05)。与对照组或Lv-NC组比较,Lv-PHLPP1组Eca109细胞中PHLPP1 mRNA和蛋白表达水平均升高(P <0. 05),细胞的增殖率、黏附率、侵袭和迁移细胞数均降低,细胞中PCNA、MMP1、MMP9、N-cadherin蛋白表达水平降低,E-cadherin蛋白表达水平升高。结论:PHLPP1在食管癌细胞中表达下调,上调其表达可以抑制食管癌细胞的增殖、黏附、侵袭和迁移能力。

Aim:To study the effects of PHLPP1 on proliferation adhesion,invasion and migration of esophageal cancer cells.Methods:The expressions of PHLPP1 in esophageal carcinoma cells Eca109,EC9706,KYSE510 and normal esophageal epithelial cells HEEC were detected by qRT-PCR and Western blot.Esophageal cancer cells were divided into three groups,the control group was not treated,the Lv-NC group infected with control recombinant lentivirus,and the Lv-PHLPP1 group infected with PHLPP1 overexpressed recombinant lentivirus.The expression of PHLPP1 was detected by qRT-PCR and Western blot,MTT assay was used to determine cell proliferation and adhesion,the cell invasion and migration ability was detected by Transwell chamber,Western blot was used to detect the expressions of PCNA,MMP1,MMP9,N-cadherin and E-cadherin in the cells.Results:The expression level of PHLPP1 in Eca109,EC9706 and KYSE510 cells was significantly lower than that in HEEC cells,and the expression level of PHLPP1 in Eca109 cells was the lowest(P<0.05).PHLPP1 mRNA and protein levels in Lv-PHLPP1 group were all elevated.The proliferation,adhesion,invasion and migration of Eca109 cells increased,the levels of PCNA,MMP1,MMP9 and N-cadherin decreased,while the level of E-cadherin protein increased in Lv-PHLPP1 group(P<0.05).Conclusion:The expression of PHLPP1 is down-regulated in esophageal cancer cells,and up-regulation of its expression can inhibit the proliferation,adhesion,invasion and migration of esophageal cancer cells.