体外沉默异戊二烯化酶二牛龙牛儿基转移酶Ⅰ对舌鳞状细胞癌增殖的影响

doi:10.7518/hxkq.2017.04.006

华西口腔医学杂志 ›› 2017, Vol. 35 ›› Issue (4): 373-378.doi: 10.7518/hxkq.2017.04.006

体外沉默异戊二烯化酶二牛龙牛儿基转移酶Ⅰ对舌鳞状细胞癌增殖的影响

王莹¹(), 王奇民², 李敬华³, 韩金宏^2,⁴, 王莉莉³, 巢晨⁵, 周建华², 童磊², 鲁旭飞^2,⁶, 周元¹, 廖奕翔², 何宗轩², 李宁⁷, 曹蕾⁷, 刘文君⁸, 陈正岗^2,⁹()

1.潍坊医学院口腔医学院,潍坊 261021
2.青岛市市立医院口腔颌面外科
3.中心实验室,青岛 266071
4.烟台市口腔医院口腔颌面外科,烟台 264008
5.南京医科大学青岛临床学院普外科,青岛 266071
6.即墨市普东卫生院口腔科,青岛 266234
7.大连医科大学研究生院,大连 116044
8.青岛市市立医院耳鼻喉科,青岛 266071
9.上海交通大学医学院附属第九人民医院口腔颌面外科,上海 200011

收稿日期:2016-10-11 修回日期:2017-04-09 出版日期:2017-08-01 发布日期:2017-08-01
作者简介:
王莹,硕士,E-mail:469055264@qq.com
基金资助:
国家自然科学基金（81372908）;青岛市卫计委计划项目（2014-WJZD009,2013-WSZD011）

Effects of geranylgeranyltransferaseⅠsilencing on the proliferation of tongue squamous cancer cells

Ying Wang¹(), Qimin Wang², Jinghua Li³, Jinhong Han^2,⁴, Lili Wang³, Chen Chao⁵, Jianhua Zhou², Lei Tong², Xufei Lu^2,⁶, Yuan Zhou¹, Yixiang Liao², Zongxuan He², Ning Li⁷, Lei Cao⁷, Wenjun Liu⁸, Zhenggang Chen^2,⁹()

1. College of Stomatology, Weifang Medical University, Weifang 261021, China
2. Dept. of Oral and Maxillofacial Surgery, Qingdao Municipal Hospital, Qingdao 266071, China
3. Central Lab, Qingdao Municipal Hospital, Qingdao 266071, China
4. Dept. of Oral and Maxillofacial Surgery, YantaiStomatological Hospital, Yantai 264008, China
5. Dept. of Surgery, Qingdao Clinical Hospital Affiliated to Nanjing Medical University, Qingdao 266071, China
6. Dept. of Stomatology, Pudong Hospital of Jimo City, Qingdao 266234, China
7. Postgraduate School, Dalian Medical University, Dalian 116044, China
8. Dept. of Otorhinolaryngology, Qingdao Municipal Hospital, Qingdao 266071, China
9. Dept. of Oral and Maxillofacial Surgery, Shanghai Ninth People’s Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai 200011, China

Received:2016-10-11 Revised:2017-04-09 Online:2017-08-01 Published:2017-08-01
Supported by:
The National Natural Science Foundation of China (81372908);Project of Qingdao Municipal Health and Family Planning Commission (2014-WJZD009, 2013-WSZD011)

摘要/Abstract

摘要：

目的研究异戊二烯化酶二牛龙牛儿基转移酶Ⅰ（GGTase-Ⅰ）在舌鳞状细胞癌增殖中的作用。方法登录Genebank确定人GGTase-Ⅰ基因序列,设计3条小干扰RNA（siRNA）,并将siRNA转染至舌癌细胞Cal-27（GGTase-ⅠsiRNA组）。设立空白对照组（只加入转染试剂,不加入siRNA）和阴性对照组（NC-siRNA）。采用实时定量聚合酶链反应和蛋白质免疫检测转染后各组细胞GGTase-Ⅰ、RhoA的mRNA和蛋白表达;蛋白质免疫检测转染48 h后Cyclin D1、p21的表达变化;细胞增殖活性检测试剂盒和流式细胞术检测细胞的增殖活性和细胞周期变化。结果与阴性对照组和空白对照组相比,GGTase-Ⅰ siRNA 组细胞的GGTase-Ⅰ的mRNA和蛋白表达下降（P<0.05）,RhoA的mRNA和蛋白表达无明显改变（P>0.05）;Cyclin D1的表达下降,p21表达升高,细胞的增殖活性下降,细胞周期发生改变（P<0.05）。结论 GGTase-Ⅰ siRNA能抑制舌鳞状细胞癌细胞中GGTase-Ⅰ的表达,抑制细胞增殖,提示GGTase-Ⅰ在舌鳞状细胞癌增殖中可能发挥重要作用。

关键词: 异戊二烯化酶二牛龙牛儿基转移酶Ⅰ, RhoA, 舌鳞状细胞癌, 增殖, Cyclin D1, p21

Abstract:

Objective This study aims to investigate the effect of geranylgeranyltransferaseⅠ (GGTase-Ⅰ) on the proliferation and growth of tongue squamous cancer cells. Methods Three small interfering RNAs (siRNAs) were designed on the basis of the GGTase-Ⅰ sequence in GeneBank. These siRNAs were then transfected into tongue squamous cancer cells Cal-27. The mRNA and protein expression of GGTase-Ⅰ and RhoA were examined by real-time quantitative polymerase chain reaction and Western blotting, respectively. The expression of Cyclin D1 and p21 were examined by Western blotting. The proliferation and growth ability were analyzed by cell counting kit-8 assay and flow cytometry. Results The mRNA and protein expression of GGTase-Ⅰ in Cal-27 was reduced significantly after the GGTase-Ⅰ siRNAs were transfected (P<0.05). No significant difference in RhoA mRNA and protein expression was detected (P>0.05). Cyclin D1 expression decreased, whereas p21 expression increased significantly. The cell cycle was altered, and the growth-proliferative activity was inhibited (P<0.05). Conclusion GGTase-Ⅰ siRNA can inhibit the expression of GGTase-Ⅰ and the proliferative activity of tongue squamous cancer cells. GGTase-Ⅰ may be a potential target for gene therapy in tongue squamous cell cancer.

Key words: geranylgeranyltransferaseⅠ, RhoA, tongue squamous cancer, proliferation, Cyclin D1, p21

中图分类号:

R739.8

王莹, 王奇民, 李敬华, 韩金宏, 王莉莉, 巢晨, 周建华, 童磊, 鲁旭飞, 周元, 廖奕翔, 何宗轩, 李宁, 曹蕾, 刘文君, 陈正岗. 体外沉默异戊二烯化酶二牛龙牛儿基转移酶Ⅰ对舌鳞状细胞癌增殖的影响[J]. 华西口腔医学杂志, 2017, 35(4): 373-378.

Ying Wang, Qimin Wang, Jinghua Li, Jinhong Han, Lili Wang, Chen Chao, Jianhua Zhou, Lei Tong, Xufei Lu, Yuan Zhou, Yixiang Liao, Zongxuan He, Ning Li, Lei Cao, Wenjun Liu, Zhenggang Chen. Effects of geranylgeranyltransferaseⅠsilencing on the proliferation of tongue squamous cancer cells[J]. West China Journal of Stomatology, 2017, 35(4): 373-378.

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siRNA	产品编号	产品名称	靶序列
GGTase-ⅠsiRNA 1	siG000005229A	si-h-PGGT1B_001	GGAACAGCTCATCCTTATG
GGTase-ⅠsiRNA 2	siG000005229B	si-h-PGGT1B_002	CGACTTAAGCCGAGTAAAT
GGTase-ⅠsiRNA 3	siG000005229C	si-h-PGGT1B_003	GACTTCTGAACGCCTTCTA

体外沉默异戊二烯化酶二牛龙牛儿基转移酶Ⅰ对舌鳞状细胞癌增殖的影响

Effects of geranylgeranyltransferaseⅠsilencing on the proliferation of tongue squamous cancer cells

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