SPOCK1通过mTOR-s6 K信号通路诱导肺癌细胞增殖、侵袭和迁移

摘要

目的:探讨SPOCK1在肺癌组织的表达及对肺癌细胞增殖、侵袭和迁移的影响.方法:肺癌细胞株A549、H3255按照转染类型分为RNA干扰组和对照组(NC组),采用小干扰RNA(siRNA)技术抑制肺癌A549、H3255细胞SPOCK1的表达,MTT增殖实验检测细胞增殖能力,集落形成实验检测细胞集落形成能力,划痕实验检测细胞迁移能力,Transwell实验检测细胞侵袭能力,RT-PCR和Western blotting检测mTOR-s6K信号通路相关mRNA和蛋白的表达水平.结果:在肺癌A549、H3255细胞中,siSPOCK1组细胞增殖率、集落形成数、侵袭细胞数、划痕愈合率明显低于NC组(P<0.05).在SPOCK1敲除的A549细胞中,siSPOCK1组和NC组G0/G1期细胞数分别为66.5％和54.3％,两组间比较差异具有统计学意义(P<0.05);在H3255细胞中,siSPOCK1组和NC组G0/G1期细胞数分别为70.2％和55.7％,两组间比较差异具有统计学意义(P<0.05).A549、H3255细胞中,与NC组比较,siSPOCK1组mTOR、s6K mRNA表达水平明显下降,组间比较差异具有统计学意义(P<0.05);siSPOCK1组p-mTOR、p-s6K蛋白相对表达水平明显低于NC组,两组间比较差异有统计学意义(P<0.05).结论:降低SPOCK1表达能够抑制肺癌细胞增殖、侵袭和迁移能力,其作用机制可能与mTOR-s6 K信号通路有关.%Objective:To explore the expression of SPOCK 1 in lung cancer tissue and its effects on prolifetion , invasion and migration .Methods:Lung cancer cell lines A 549 , H3255 were divided into interfere group and con-trol group (NC group) according to interfection method, the expression of SPOCK1 in lung cancer cell line A549 and H3255 were inhibited by siRNA , the proliferation of cells were measured by MTT assay , the colony formation capacity were detected by colony formation assay , the migration capacity were mearsured by wound heal assay , the cell invasion capacity were mearsured by Transwelll assay , the expression of mRNA and protein related to mTOR-s6K pathway were measured by RT-PCR and Western blotting.Results: In A549 and H3255 cell lines, the cell proliferation rate , number of colonies , number of invasion cells , wound healing rates in siSPOCK 1 group were sig-nificantly less than those in NC group , the difference were statistically significant (P<0.05).In SPOCK1 knock-down A549 cells, the G0/G1 phases cell were 66.5％ and 54.3％ in siSPOCK1 and NC group, respectively, the difference between the two groups was statistically significant (P<0.05).And in H3255 cells, the G0/G1 phases cell were 70 .2％and 55 .7％in siSPOCK1 and NC group , respectively , the difference between the two groups was statistically significant (P<0.05).In A549 and H3255 cell linse, the expression of mTOR, s6K mRNA in siS-POCK1 group was statistically decreased when compared to NC group , the difference between the two groups was statistically significant (P<0.05).The expression of p-mTOR, p-s6K protein in siSPOCK1 group were statistically decreased when compared to NC group , the difference between the two groups was statistically significant ( P <0 .05 ) .Conclusion:Inhibition the expression of SPOCK 1 can reduce the proliferation , invasion and migration ca-pability of lung cancer cells , and its mechanism may be related to mTOR-s6K signaling pathway .