组织激肽释放酶7高表达前列腺癌PC-3单克隆细胞株的构建及其侵袭性研究  

Research of establishment of a human prostatic carcinoma cell line PC-3 stably overexpressing Kallikrein 7 and tumor invasion

作　　者：吕文鑫[1] 莫曾南[2,3] 杨小丽[2,4] LV Wenxin;MO Zengnan;YANG Xiaoli(Department of Urology,Liuzhou Worker's Hospital,Guangxi Zhuang Autonomous Region(The Fourth Affiliated Hospital of Guangxi Medical University),Liuzhou 545005)

机构地区：[1]广西壮族自治区柳州市工人医院(广西医科大学第四附属医院)泌尿外科,柳州545005 [2]广西医科大学第一附属医院,南宁530021 [3]广西医科大学基因组与个体化医学研究中心,南宁530021 [4]广西医科大学医学科学实验中心,南宁530021

出　　处：《陕西医学杂志》2020年第5期527-530,共4页Shaanxi Medical Journal

基　　金：国家自然科学基金资助项目(81060214)。

摘　　要：目的:构建重组人组织激肽释放酶7(KLK7)表达载体,建立稳定过表达KLK7的前列腺癌细胞株PC-3,并研究其细胞侵袭性。方法:将扩增后的KLK7cDNA克隆到表达载体pcDNA3.1上;使用限制性内切酶酶切后,DNA测序验证。然后,通过脂质体法转染人前列腺癌细胞株PC-3;选用G418进行筛选,并扩大培养每个单克隆细胞,进而建立稳定转染后的KLK7单克隆细胞株PC-3;Western blot检测目的蛋白的表达。通过细胞侵袭性试验了解其细胞侵袭性。结果:①通过酶切鉴定、DNA测序分析并证明,成功构建pcDNA3.1-KLK7表达载体;②成功获得稳定过表达KLK7的前列腺癌单克隆细胞株PC-3;③Western blot结果显示:经过pcDNA3.1-KLK7转染后的细胞与未转染的细胞表达量比较有统计学差异(P<0.01);④细胞侵袭性试验结果显示:转染后的细胞侵袭性高于未转染的细胞。结论:成功的建立pcDNA3.1-KLK7表达载体及稳定过表达KLK7的前列腺癌PC-3单克隆细胞株,同时证明其细胞侵袭性,为研究KLK7在前列腺癌发生、发展中的作用奠定了基础。Objective:To construct the expression vector containing Kallikrein 7(KLK7),establish stable cell line with high expression of KLK7,and study tumor invasion.Methods:Amplified KLK7 cDNA was cloned into expression vector pcDNA3.1 and sequenced after restriction endonuclease digestion.Then,the human prostatic carcinoma cell line PC-3 was transfected by liposome method.G418 was selected for screening,and each monoclonal cell was expanded to culture,and then the stable transfected KLK7 monoclonal cell line PC-3 was established.Western blot was used to detect the expression of the target protein.The cellular invasiveness was studied by tumor invasion assay.Results:The pcDNA3.1-KLK7 expression vector was successfully constructed by enzyme digestion and DNA sequencing analysis.The prostate cancer cell line PC-3 with stable overexpression of KLK7 was successfully obtained.The results of Western blot showed that the expression of pcDNA3.1-KLK7 transfected cells was significantly different from that of untransfected cells(P<0.01).The results of tumor invasion assay showed that the invasiveness of transfected cells was higher than that of untransfected cells.Conclusion:The construction of the expression vector pcDNA3.1-KLK7 and the establishment of the human prostatic carcinoma cell line PC-3 monoclonal cell line overexpressed KLK7 stably is successful,and the invasiveness is proved.It lays a foundation for studying the role of KLK7 in the occurrence and development of prostatic carcinoma.

关 键 词：组织激肽释放酶7 前列腺癌 稳定细胞株 表达载体 细胞侵袭性 

分 类 号：R737.25[医药卫生—肿瘤]