摘要
目的探讨组蛋白去甲基化酶JMJD1A对宫颈癌细胞增殖的作用。方法应用免疫组织化学和Western blot方法检测JMJD1A蛋白在33例人正常宫颈、48例宫颈鳞状细胞癌组织以及宫颈癌细胞系(SiHa、HeLa、CaSki和C-33A)中的表达情况。采用JMJD1A-shRNA质粒稳定干扰HeLa和SiHa细胞系中JMJD1A基因的表达。采用MTT法分析JMJD1A干扰后的宫颈癌细胞增殖的变化。裸鼠成瘤实验分析JMJD1A下调后对裸鼠移植瘤瘤体生长、体积及质量的变化。结果与正常宫颈组织相比,宫颈癌组织中JMJD1A蛋白水平显著升高(P<0.001)。Western blot结果显示JMJD1A蛋白高表达于各宫颈癌细胞SiHa、HeLa、CaSki和C-33A中。JMJD1A-shRNA质粒能显著抑制JMJD1A基因表达。JMJD1A基因下调后,宫颈癌细胞生长速度及活性明显下降,且具有时间依赖效应(P<0.01)。与对照组(HeLa-Ctrl/SiHa-Ctrl)相比较,干扰组(HeLa-shJMJD1A/SiHa-shJMJD1A)裸鼠移植瘤生长明显减慢,且移植瘤的平均质量均显著减小(P<0.01)。结论JMJD1A表达与宫颈癌发生发展呈正相关,可显著促进宫颈癌细胞增殖能力,提示JMJD1A可能作为一个癌基因发挥作用。
Objective To investigate the effect of histone demethylase JMJD1A on the proliferation of cervical cancer cells.Methods Immunohistochemistry and Western blot were used to determine JMJD1A protein in 33 normal cervixes,48 cervical squamous cell carcinoma tissues and cervical cancer cell lines(SiHa,HeLa,CaSki,C-33A).JMJD1A-shRNA interference plasmid was constructed and used to stably downregulate JMJD1A expression in HeLa and SiHa cell lines.MTT assay and nude mice xenograft ananlysis were used to determine the effect of JMJD1A oncervical cancer cell proliferation in vitro and the growth,volume and mass of xenografts in HeLa and SiHa cells after downregulating JMJD1A.Results JMJD1A expression level was found to be significantly elevated in cervical cancer tissues compared to normal cervixes(P<0.001).Western blot results showed that JMJD1A protein was highly expressed in cervical cancer cells(SiHa,HeLa,CaSki and C-33A).The expression of JMJD1A was significantly inhibited by JMJD1A-shRNA plasmid.The cell growth and viability were significantly decreased after JMJD1A downregulation in vitro in a time-dependent manner(P<0.01).Compared with control groups(HeLa-Ctrl/SiHa-Ctrl),the downregulation of JMJD1A in HeLa and SiHa cells significantly attenuated the growth and mass of xenografts in nude mice in vivo(P<0.01).Conclusion JMJD1A expression is positively correlated with cervical carcinogenesis,and could significantly promote cell proliferation,suggesting that JMJD1A may play a role as an oncogene in cervical cancer.
作者
吴晓玲
万秋园
杨阳
樊江波
WU Xiaoling;WAN Qiuyuan;YANG Yang;FAN Jiangbo(Department of Obstetrics and Gynecology,Second Affiliated Hospital of Xi’an Jiaotong University,Xi’an 710004,China)
出处
《山西医科大学学报》
CAS
2019年第12期1655-1660,共6页
Journal of Shanxi Medical University
基金
国家自然科学基金资助项目(81702578)
陕西省重点研发计划项目(2017SF-082)
关键词
JMJD1A
宫颈癌
细胞增殖
移植瘤
JMJD1A
cervical cancer
cell proliferation
tumor xenograft
