人生长激素释放激素受体剪接变异体1型对人肝癌细胞HepG2增殖的影响

doi:10.13481/j.1671-587x.20170303

吉林大学学报(医学版) ›› 2017, Vol. 43 ›› Issue (03): 474-478.doi: 10.13481/j.1671-587x.20170303

人生长激素释放激素受体剪接变异体1型对人肝癌细胞HepG2增殖的影响

林玉茵¹, 陈文生¹, 郭晓兰¹, 谭茵¹, 贺小松², 戴建威¹

1. 广州医科大学生命科学学院生物技术系, 广东广州 511436;
2. 广州医科大学基础学院人体解剖学教研室, 广东广州 511436

收稿日期:2016-08-12 出版日期:2017-05-28 发布日期:2017-06-01
通讯作者: 戴建威,副教授,硕士研究生导师(Tel:020-37103208,E-mail:daijw@gzhmu.edu.cn) E-mail:daijw@gzhmu.edu.cn
作者简介:林玉茵(1993-),女,广东省广州市人,在读医学硕士,主要从事基础医学方面的研究。
基金资助:
国家自然科学基金资助课题（81501014）；广东省科技厅自然科学基金资助课题（2016A030313601）；广东省广州市科技局科技计划项目资助课题（201510010076）

Effect of human growth hormone releasing hormone receptor splice variant type 1 on proliferation of human liver cancer HepG2 cells

LIN Yuyin¹, CHEN Wensheng¹, GUO Xiaolan¹, TAN Yin¹, HE Xiaosong², DAI Jianwei¹

1. Department of Biotechnology, School of Life Science, Guangzhou Medical University, Guangzhou 511436, China;
2. Department of Human Anatomy, School of Basic Medical Sciences, Guangzhou Medical University, Guangzhou 511436, China

Received:2016-08-12 Online:2017-05-28 Published:2017-06-01

摘要/Abstract

摘要： 目的：探讨人生长激素释放激素受体剪接变异体1型（GHRHR SV1）对肝癌细胞系HepG2增殖的影响，阐明GHRHR SV1对人肿瘤细胞的促增殖作用。方法：将GHRHR SV1真核表达载体导入HepG2细胞建立HepG2-SV1细胞系。设计对照组（野生型HepG2）、空载质粒组（HepG2-pCDNA3.0细胞系）和干扰组（HepG2-SV1细胞系）。采用PCR和Westernblotting法鉴定HepG2-SV1细胞系，CCK-8法检测各组细胞的增殖率，克隆形成实验检测各组细胞的单克隆形成率，细胞划痕实验检测细胞的迁移率。结果：PCR与Westernblotting法检测，构建的HepG2-SV1细胞系能稳定表达GHRHR SV1；CCK-8法检测，干扰组HepG2-SV1细胞系的增殖率高于空载质粒组HepG2-pcDNA3.0细胞系（P<0.05）；克隆形成实验，干扰组HepG2-SV1细胞系的单克隆形成率约为空载质粒组HepG2-pcDNA3.0细胞系的3.5倍（P<0.05）；细胞划痕实验，所构建的干扰组HepG2-SV1细胞系的迁移率高于空载质粒组HepG2-pcDNA3.0细胞系（P<0.05）。结论：GHRHR SV1能促进HepG2细胞的增殖。

关键词: 人生长激素释放激素受体剪接变异体1型, 细胞增殖, 迁移率, HepG2细胞, 克隆形成率

Abstract: Objective: To investigate the effect of human growth hormone releasing hormone receptor splice variant type 1 (GHRHR SV1) on the proliferation of human liver cancer HepG2 cells, and to clarify the proliferation effect of GHRHR SV1 on the human cancer cells. Methods: The GHRHR SV1 plasmids were transfected into the human HepG2 cells to construct the HepG2-SV1 cell line. HepG2 group(HepG2 cells),HepG2-empty group(HepG2-pcDNA3.0 cell line) and HepG2-SV1 group(HepG2-SV1 cells) were set up. PCR and Western blotting methods were used to identify the HepG2-SV1 cell line;CCK-8 method was used to detect prolifernation rate of cells;colony formation assay was used to detect the colony formation rate of cells;cell wound healing assay was used to evaluate the migration rate of cells. Results: The PCR and Western blotting results showed the HepG2-SV1 cell line expressed GHRHR SV1 steadily.The CCK-8 results showed that the proliferation rate of the HepG2-SV1 cells in HepG2-SV1 group was higher than that of the HepG2-pcDNA3.0 cells in HepG2-empty group(P<0.05).The colony formation assay results showed that the colony formation rate of HepG2-SV1 cells in HepG2-SV1 group was 3.5 times higher than that of the HepG2-pcDNA3.0 cells in HepG2-empty group(P<0.05).The cell wound scratch assay results showed that the migration rate of the HepG2-SV1 cells in HepG2-SV1 group was higher than that of the HepG2-pcDNA3.0 cells in HepG2-empty group(P<0.05). Conclusion: GHRHR SV1 could increase the proliferation of HepG2 cells.

Key words: HepG2 cells, cell proliferation, colony formation rate, migration rate, human growth hormone releasing hormone receptor splice variant type 1

中图分类号:

R735.7

林玉茵, 陈文生, 郭晓兰, 谭茵, 贺小松, 戴建威. 人生长激素释放激素受体剪接变异体1型对人肝癌细胞HepG2增殖的影响[J]. 吉林大学学报(医学版), 2017, 43(03): 474-478.

LIN Yuyin, CHEN Wensheng, GUO Xiaolan, TAN Yin, HE Xiaosong, DAI Jianwei. Effect of human growth hormone releasing hormone receptor splice variant type 1 on proliferation of human liver cancer HepG2 cells[J]. Journal of Jilin University Medicine Edition, 2017, 43(03): 474-478.

参考文献

[1] Hohla F,Moder A,Mayrhauser U,et al.Differential expression of GHRH receptor and its splice variant 1 in human normal and malignant mucosa of the oesophagus and colon[J].Int J Oncol,2008,33(1):137-143.
[2] Papadia A,Schally AV,Halmos G,et al.Growth hormone-releasing hormone antagonists inhibit growth of human ovarian cancer[J].Horm Metab Res,2011,43(11):816-820.
[3] Dioufa M,Farmaki E,Schally AV,et al.Growth hormone-releasing hormone receptor splice variant 1 is frequently expressed in oral squamous cell carcinomas[J].Horm Cancer,2012,3(4):172-180.
[4] Chatzistamou I,Volakaki A,Schally AV,et al.Expression of growth hormone-releasing hormone receptor splice variant 1 in primary human melanomas[J].Regul Pept,2008,147(1-3):33-36.
[5] Mezey G,Treszl A,Schally AV,et al.Prognosis in human glioblastoma based on expression of ligand growth hormone-releasing hormone,pituitary-type growth hormone-releasing hormone receptor,its splicing variant receptors,EGF receptor and PTEN genes[J].J Cancer Res Clin Oncol,2014,140(10):1641-1649.
[6] 郭晓兰,谭茵,陈文生,等.GHRHR-SV1抗体的制备及其蛋白在黑色素瘤细胞中的表达[J].吉林大学学报:医学版,2016,42(6):1237-1242.
[7] 田力,张蕾,李伟伟,等.人生长激素释放激素变异受体1型基因绿色荧光蛋白真核表达载体1的构建及体外表达[J].江西医学学报,2009,49(3):15-20.
[8] 汪期明,黄勇.生长激素释放激素受体剪接变异体1在子宫内膜异位症中的表达[J].中国妇幼保健,2012,27(28):4441-4444.
[9] 沈烨,胡吉,张玉林,等.生长激素释放激素在垂体生长激素瘤中的作用[J].复旦学报:医学版,2006,33(4):463-466.
[10] 黄勇,汪期明.生长激素释放激素拮抗剂对人异位子宫内膜抑制增殖作用与cAMP信号通路相关[J].医学研究杂志,2013(5),103-106.
[11] Szalontay L,Schally AV,Popovics P,et al.Novel GHRH antagonists suppress the growth of human malignant melanoma by restoring nuclear p27 function[J].Cell Cycle,2014,13(17):2790-2797.
[12] Munoz-Moreno L,Arenas MI,Schally AV,et al.Inhibitory effects of antagonists of growth hormone-releasing hormone on growth and invasiveness of PC3 human prostate cancer[J].Int J Cancer,2013,132(4):755-765.
[13] Kovacs M,Schally AV,Hohla F,et al.A correlation of endocrine and anticancer effects of some antagonists of GHRH[J].Peptides,2010,31(10):1839-1846.
[14] Munoz-Moreno L, Arenas MI, Carmena MJ, et al.Growth hormone-releasing hormone antagonists abolish the transactivation of human epidermal growth factor receptors in advanced prostate cancer models[J].Invest New Drugs,2014,32(5):871-882.
[15] Rick FG,Schally AV,Szalontay L,et al.Antagonists of growth hormone-releasing hormone inhibit growth of androgen-independent prostate cancer through inactivation of ERK and Akt kinases[J].Proc Natl Acad Sci USA,2012,109(5):1655-1660.
[16] Catanuto P,Tashiro J,Rick F,et al.Expression of receptors for pituitary-type growth hormone-releasing hormone (pGHRH-R) in human papillary thyroid cancer cells:effects of GHRH antagonists on matrix metalloproteinase-2[J].Horm Cancer,2015,6(2/3):100-106.

人生长激素释放激素受体剪接变异体1型对人肝癌细胞HepG2增殖的影响

Effect of human growth hormone releasing hormone receptor splice variant type 1 on proliferation of human liver cancer HepG2 cells

RichHTML

PDF (PC)

赞

可视化

摘要/Abstract

引用本文

使用本文

参考文献

相关文章 15

Metrics

本文评价

推荐阅读 0

[1]	朱德强, 陈学军. 白花丹素对肝癌索拉菲尼耐药细胞HepG2R增殖和凋亡的影响及其机制[J]. 吉林大学学报(医学版), 2018, 44(06): 1223-1229.
[2]	王廷刚, 薛峰, 李宇, 牛兆健, 王野. miR-26a靶向HMGA1基因对结肠癌细胞生长、侵袭和迁移的影响[J]. 吉林大学学报(医学版), 2018, 44(06): 1205-1211.
[3]	阎慧, 马淑飞, 段明华, 闫玉礼, 潘新, 李海清, 周长龙, 赵天倚. 京尼平对胃癌SGC 7901细胞体外增殖、侵袭能力和凋亡的影响及其机制[J]. 吉林大学学报(医学版), 2018, 44(05): 924-928.
[4]	邢树刚, 张丽红, 金明华, 李百慧, 武则馨, 房波, 李伟. 杨梅黄酮对小鼠脑胶质瘤GL261细胞增殖和凋亡的影响[J]. 吉林大学学报(医学版), 2018, 44(05): 955-961.
[5]	袁虎勤, 李强. 水飞蓟素对人胃癌SGC 7901细胞增殖、迁移和侵袭的抑制作用及其机制[J]. 吉林大学学报(医学版), 2018, 44(05): 988-993.
[6]	刘定坤, 杨楠, 金巨楼, 孙梦洁, 王倩, 吴佳, 刘志辉. 牙髓干细胞增殖、分化和迁移的研究进展[J]. 吉林大学学报(医学版), 2018, 44(05): 1100-1104.
[7]	李鑫, 马云飞, 唐庚, 韦麒, 纪红池, 田嘉安, 申延男, 王志成. 线粒体靶向KillerRed诱导的ROS增强辐射对HeLa细胞的增殖抑制作用[J]. 吉林大学学报(医学版), 2018, 44(04): 718-723.
[8]	朱效慧, 孟琴, 冯世燕. TP53在不明原因复发性流产患者绒毛组织中的表达及其对人滋养层细胞生长的影响[J]. 吉林大学学报(医学版), 2018, 44(04): 796-800.
[9]	李天柱, 王加茹, 张俊毅, 王洪权, 史铁伟, 周静, 白春英, 金成浩. ELK-3蛋白在肝癌组织中的表达及其对肝癌HepG2细胞迁移和侵袭能力的影响[J]. 吉林大学学报(医学版), 2018, 44(04): 736-740.
[10]	王淼, 母润红, 李明成, 李洪杰. RNAi沉默survivin和HIF-1α基因对胃癌BGC-823细胞体外增殖和凋亡的影响[J]. 吉林大学学报(医学版), 2018, 44(04): 753-758.
[11]	吕鹏, 宁明杰, 邵玉, 张璐妮, 唐英, 王南博, 陈飞儿, 齐玲. 野黄芩苷对人脑胶质瘤U87细胞的增殖抑制作用及其机制[J]. 吉林大学学报(医学版), 2018, 44(03): 466-470.
[12]	杨文延, 刘强, 孙志娟, 杜利清, 徐畅, 王彦, 柳杨, 王芹. 褪黑素对人非小细胞肺癌H1299细胞辐射敏感性的影响[J]. 吉林大学学报(医学版), 2018, 44(03): 532-536.
[13]	张蕾, 张琨, 王德利, 任小红, 崔娜, 刘洋, 赵庭勋. 桦褐孔菌皂苷的优化提取及其对HepG2细胞脂肪堆积的清除作用[J]. 吉林大学学报(医学版), 2018, 44(03): 526-531.
[14]	周刘祥, 林琳, 胡鑫敏, 王晓春. 内分泌腺性血管内皮生长因子对结肠癌LoVo细胞增殖和迁移的促进作用[J]. 吉林大学学报(医学版), 2018, 44(03): 516-520.
[15]	上官梦原, 赵菁, 杨艳荣, 张佳悦, 陈俊宇, 赵淑华. 五味子脂A联合卡铂对人卵巢癌Skov3细胞增殖、迁移和侵袭的抑制作用及其机制[J]. 吉林大学学报(医学版), 2018, 44(02): 292-298.