目的:应用慢病毒载体shRNA对PC3细胞中的人趋化素样因子超家族成员3(CKLF-like MARVEL transmembrane domain containing 3,CMTM3)进行敲减,以探究CMTM3对前列腺癌细胞系生物学行为改变的影响。方法:研究分为两组进行,其中sh N为未敲减CMTM3的PC3细胞系对照组,sh393为敲减CMTM3的PC3细胞系实验组。运用Western blot检测PC3细胞系慢病毒shRNA敲减CMTM3后的表达,Transwell与细胞划痕实验检测敲减CMTM3对PC3细胞系迁移能力的影响,Matrigel实验检测敲减CMTM3对PC3细胞系侵袭能力的影响。结果:前列腺癌细胞系PC3经慢病毒敲减CMTM3后,sh393组CMTM3表达水平明显低于sh N组(0.004 0±0.000 4 vs.0.490 0±0.055 7,P〈0.001),且sh393组侵袭(248.6±4.5 vs.113.0±3.3)、迁移(203.6±1.9 vs.103.0±1.2)及迁移愈合能力(95.0±2.9 vs.33.0±1.5)均明显强于sh N组(P〈0.001)。结论:敲减CMTM3可以影响PC3细胞的迁移与侵袭能力,CMTM3下调与PC3细胞系肿瘤生物学特性的改变可能具有负相关性。
Objective:To investigate the change of biological characteristics after stable knockdown of CKLF-like MARVEL transmembrane domain containing 3 (CMTM3) expression in PC3 by lentivirus shRNA and to reveal new therapeutic targets. Methods: The research includes two groups: sh393 is the experimental group in which CMTM3 is knocked down in PC3 cell line; shN is the control group in which CMTM3 is negatively knocked down. The expression of CMTM3 was detected by Western blot. The migration ability of PC3 after stable knockdown was detected by Transwell and Wound healing assay. The invasion ability of PC3 was detected by Matrigel assay. Results were obtained from at least three indivi- dual experiments. Results: The expression of CMTM3 in sh393 group is significant lower than shN group (0. 004 0 ±0. 000 4 vs. 0. 490 0 ±0. 055 7, P 〈0. 001 ) detected by Western blot. It also had statistical significance in Matrigel assays (248.6 ± 4.5 vs. 113.0 ± 3.3 ), Transwell (203.6 ±1.9 vs. 103.0 ± 1.2) and Wound healing assays (95.0 ±2.9 vs. 33.0 ± 1.5 ) that knockdown of CMTM3 promoted migration, and invasion of PC3 cells in vitro (P 〈 0. 001 ). Conclusion: Negative correlation exists between the stable knockdown of CMTM3 and change of biological characteristics in PC3 cells, and knocking down CMTM3 affects migration, and invasion ability in PC3 cells.