[1]吕文鑫,莫曾南,杨小丽△.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*[J].陕西医学杂志,2020,49(5):527-530.
　LV Wenxin,MO Zengnan,YANG Xiaoli,et al.Research of establishment of a human prostatic carcinoma cell line PC-3 stably overexpressing Kallikrein 7 and tumor invasion[J].,2020,49(5):527-530.
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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*
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《陕西医学杂志》[ISSN:1000-7377/CN:61-1281/TN]

卷:
49
期数:
2020年5期
页码:
527-530
栏目:
基础研究
出版日期:
2020-05-05

文章信息/Info

Title:
Research of establishment of a human prostatic carcinoma cell line PC-3 stably overexpressing Kallikrein 7 and tumor invasion
文章编号:
DOI:10.3969/j.issn.10007377.2020.05.004
作者:
吕文鑫1; 莫曾南2; 3 ; 杨小丽2; 4△
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)
Author(s):
LV Wenxin; MO Zengnan; YANG Xiaoli; et al.
Department of Urology,Liuzhou Worker's Hospital,Guangxi Zhuang Autonomous Region(The Fourth Affiliated Hospital of Guangxi Medical University)(Liuzhou 545005)
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
Keywords:
Kallikrein 7;  Prostate cancer;  Stable cell line;  Eukaryotic expression vector;  Cellular invasiveness
分类号:
R737.25
文献标志码:
A
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。
Abstract:
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.

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备注/Memo

备注/Memo:
*国家自然科学基金资助项目(81060214)
更新日期/Last Update: 2020-07-28