着丝粒蛋白K小干扰RNA转染对人肺癌细胞系A549增殖、迁移和侵袭能力的影响  

Effects of si-CENPK transfection on proliferation,migration,and invasion of human lung cancer A549 cells

作　　者：叶侠 周琳[1] 吴杰[1] 陶敏[1] YE Xia;ZHOU Lin;WU Jie;TAO Min(The First Affiliated Hospital of Soochow University,Suzhou 215000,China)

机构地区：[1]苏州大学附属第一医院

出　　处：《山东医药》2020年第3期1-4,共4页Shandong Medical Journal

基　　金：国家自然科学基金资助项目(81772645,81572992);江苏省高等学校自然科学研究项目(18KJB320025)

摘　　要：目的观察着丝粒蛋白K(CENPK)小干扰RNA(si-CENPK)转染对人肺癌细胞系A549增殖、迁移和侵袭能力的影响。方法体外传代培养A549细胞及正常人支气管上皮细胞系HBE,采用q-RTPCR法检测CENPK mRNA。取生长状态良好的A549细胞,随机分为3组,实验组转染si-CENPK,阴性对照组转染si-Negative Control,空白对照组不做转染;转染48 h后,收集三组细胞,采用q-RTPCR法检测细胞CENPK mRNA,CCK-8法检测转染后24、48、72、96 h OD值,细胞划痕实验检测转染后48 h细胞间距离,Transwell侵袭实验检测转染后48 h穿膜细胞数。结果 A549细胞、HBE细胞中CENPK mRNA相对表达量分别为2. 710±0. 153、1. 000,两者比较,P均<0. 05。si-CENPK实验组、si-NC阴性对照组、空白对照组CENPK mRNA相对表达量分别为0. 28±0. 06、1. 12±0. 10、1. 000,si-CENPK实验组与其他两组比较,P <0. 05。si-CENPK实验组培养24、48、72、96 h OD值分别为0. 317±0. 003、0. 512±0. 030、0. 711±0. 052、1. 012±0. 090,si-NC阴性对照组分别为0. 324±0. 004、0. 602±0. 022、0. 920±0. 043、1. 370±0. 103,空白对照组分别为0. 322±0. 005、0. 610±0. 040、0. 918±0. 054、1. 320±0. 110,si-CENPK实验组与其余两组比较,P均<0. 05。si-CENPK实验组、si-NC阴性对照组、空白对照组转染48 h细胞间距离分别为(0. 70±0. 02)、(0. 34±0. 03)、(0. 37±0. 06) cm,si-CENPK实验组、si-NC阴性对照组比较,P <0. 05。si-CENPK实验组、si-NC阴性对照组、空白对照组转染48 h穿膜细胞数分别为(89±10)、(163±12)、(168±11)个,si-CENPK实验组与其他两组比较,P均<0. 05。结论 si-CENPK转染能够抑制A549细胞增殖、迁移和侵袭能力。Objective To explore the effects of centromere protein K small interfering RNA( si-CENPK) on the proliferation,migration,and invasion of human lung cancer A549 cells. Methods Human lung cancer cell line A549,and normal bronchial epithelial cells HBE were cultured in vitro. The expression of CENPK mRNA was detected by q-RTPCR.A549 cells in good growth was randomly divided into three groups: the si-CENPK experimental group( transfected with siCENPK),si-NC negative control group( transfected with si-Negative Control),and blank control group( not transfected).After 48-hour transfection,the cells were collected,and the expression of CENPK mRNA in three groups was detected by real-time quantitative PCR. CCK-8 was applied to detect OD values at 24,48,72,96 h after 48-hour transfection,and cell Scratch assay was used to detect the distance between cells after 48-hour transfection,and Transwell invasion assay was applied to detect the number of transmembrane cells after 48-hour transfection. Results The relative expression of CENPK mRNA in A549 and HBE cells was 2. 710 ± 0. 153 and 1. 000,respectively,with statistically significant difference( P< 0. 05). The relative expression of CENPK mRNA in the si-CENPK experimental group,si-NC negative control group and blank control group was 0. 28 ± 0. 06,1. 12 ± 0. 10,and 1. 000,respectively,with statistically significant difference( P <0. 05). The OD values of si-CENPK at 24,48,72,and 96 h were 0. 317 ± 0. 003,0. 512 ± 0. 030,0. 711 ± 0. 052,and1. 012 ± 0. 090,respectively,in the si-CENPK experimental group,0. 324 ± 0. 004,0. 602 ± 0. 022,0. 920 ± 0. 043,and1. 370 ± 0. 103,respectively,in the si-NC negative control group,and 0. 322 ± 0. 005,0. 610 ± 0. 040,0. 918 ± 0. 054,1. 320 ± 0. 110,respectively,in the blank control group,with statistically significant difference( all P < 0. 05). The distance between cells in the si-CENPK experimental group,si-NC negative control group,and blank control group after 48-hour transfection was( 0. 70 ± 0. 02),( 0. 34 ± 0. 03),an

关 键 词：肺癌 着丝粒蛋白K 细胞增殖 细胞迁移 细胞侵袭 

分 类 号：R734.2[医药卫生—肿瘤]