沉默C20ORF32表达对结直肠癌LOVO细胞增殖与侵袭的影响  

Effect of C20ORF32 shRNA on proliferation and invasion of colorectal cancer LOVO cells

作　　者：乔鲁冀[1] 岳滨[1] 戴顺东[2] 张春霞[1] 

机构地区：[1]沈阳市肛肠医院肛肠外科,沈阳110002 [2]中国医科大学基础医学院病理学教研室,沈阳110122

出　　处：《解剖科学进展》2016年第1期9-12,15,共5页Progress of Anatomical Sciences

基　　金：国家自然科学基金(81401881);辽宁省科技厅自然科学基金(201402018)

摘　　要：目的探讨沉默Crk相关底物家族成员C20ORF32基因表达后,对结直肠癌细胞增殖和侵袭的影响。方法应用蛋白印迹法(Western blot)检测4种结直肠癌细胞系(LOVO、SW620、HT-29和SW480)中C20ORF32的表达状况;用装载了C20ORF32 shRNA的慢病毒颗粒,感染高表达C20ORF32的结直肠癌细胞系LOVO后,运用免疫荧光染色和蛋白质免疫印迹的方法检测其对C20ORF32基因的沉默效果,并通过MTT法和软琼脂克隆形成实验检测对细胞增殖能力的影响,用Transwell侵袭实验检测细胞的体外侵袭能力。结果与其他3株细胞系相比,C20ORF32蛋白在LOVO细胞中明显高表达(t=15.33,P<0.001)。下调C20ORF32表达后,LOVO细胞的增殖速度在24h以及48h显著降低(24hF=81.32,P<0.001;48hF=13.33,P<0.05),集落数目明显减少(F=74.21,P=0.0002),细胞侵袭能力明显减弱(F=144.38,P<0.001)。结论 C20ORF32的高表达可能与结直肠癌细胞的快速增殖与侵袭有关,参与了结直肠癌的恶性进展过程。Objective To explore the role of C20ORF32 in the proliferation and invasion of colorectal carcinoma（CRC） cells.Methods Western-blot was used to detect C20ORF32 expression in 4 kinds of CRC cell lines.The lentiviral vector was constructed to silence C20ORF32 gene in LOVO cells.Immunofluorescence staining and Western blot were used to detect the silence effect of＇C20ORF32.Then,methylthiazolyl tetrazolium（MTT） and soft agar assay were used to detect cell proliferation.Besides,cell invasive ability was measured by Transwell invasion assay.Results The expression level of C20ORF32 was significantly higher in the highly metastatic potential LOVO and SW620 cell lines than in the low metastatic HT-29 and SW480 cell lines（t = 15.33,P〈0.001）.After silencing C20ORF32 in LOVO cells at 24 and 48 hours,the cell proliferation ability（24hF=81.32,P〈0.001;48hF=13.33,P〈0.05）,number of shC20ORF32colonies（24h F=81.32,P〈0.001;48h F=13.33,P〈0.05）,invasion ability（F=144.38,P〈0.001） of LOVO cells were significantly decreased.Conclusion C200RF32 overexpression may be related to proliferation and invasion ability of LOVO cells,which involved in CRC development and progression.

关 键 词：结直肠癌LOVO细胞系 C20ORF32 增殖 侵袭 

分 类 号：R735.3[医药卫生—肿瘤]