瘦素对HTR8-SVneo细胞增殖的影响及调控机制

山东大学学报(医学版) ›› 2012, Vol. 50 ›› Issue (1): 51-56.

瘦素对HTR8-SVneo细胞增殖的影响及调控机制

程欢欢1，王华阳1，董召刚2，徐小飞3，孔北华3，曲迅1

山东大学齐鲁医院 1.临床基础研究所； 2.检验科； 3.妇产科，济南 250012

收稿日期:2011-09-16 出版日期:2012-01-10 发布日期:2012-01-10
通讯作者: 曲迅(1958- )，女，教授，博士生导师，主要从事生殖免疫的研究。 Email：quxun@sdu.edu.cn
作者简介:程欢欢(1986- )，女，硕士研究生，主要从事生殖免疫的研究。
基金资助:
国家自然科学基金资助课题(81072406，30872321)；山东省自然科学基金资助课题(Y2008C02)；山东大学研究生自主创新基金资助(yzc10133)；山东大学优秀研究生科研创新基金资助(yyx10126)。

Effect of leptin on HTR8-SVneo cell proliferation in vitro and its mechanism

CHENG Huan-huan1, WANG Hua-yang1, DONG Zhao-gang2,
XU Xiao-fei3, KONG Bei-hua3, QU Xun1

1. Institute of Basic Medical Sciences; 2. Department of Clinical Laboratory;
3. Department of Gynaecology and Obstetrics, Qilu Hospital of
Shandong University, Jinan 250012, China

Received:2011-09-16 Online:2012-01-10 Published:2012-01-10

摘要/Abstract

摘要：

目的研究瘦素(Leptin)对HTR8-SVneo滋养细胞系增殖的影响及对DNA结合和分化抑制剂(ID)分子表达水平的影响，探讨Leptin对HTR8SVneo细胞增殖作用的调节机制。方法体外培养HTR8-SVneo滋养细胞，MTT法检测Leptin不同质量浓度(0、10、50、100、250、500ng/mL)刺激后细胞增殖情况。HTR8-SVneo细胞培养体系中加入Leptin(0、100、500ng/mL)，RT-PCR检测瘦素受体(Leptin R)各亚型表达，实时定量PCR检测ID分子表达，Western Blot检测ID2表达。SiRNA干扰ID2分子后，分为Leptin(0ng/mL)、Leptin(100ng/mL)、Leptin(500ng/mL)、Leptin(0ng/mL)+ID2siRNA、Leptin(100ng/mL)+ID2siRNA、Leptin(500ng/mL)+ID2-siRNA，MTT检测Leptin对HTR8-SVneo滋养细胞增殖的影响。结果与Leptin(0ng/mL)比较，高质量浓度Leptin(100、250、500ng/mL)促进HTR8-SVneo细胞增殖(P<0.05)；HTR8-SVneo细胞表达除HuB219.2外的全部受体亚型；Leptin以剂量依赖方式促进HTR8SVneo细胞ID2分子及OB-RL表达(P<0.05)；ID2-siRNA抑制ID2表达并逆转Leptin对滋养细胞增殖的促进作用。结论 Leptin可能通过Leptin R-ID2途径促进HTR8-SVneo细胞增殖，为阐明滋养细胞增殖调控机制提供了新的基础数据。

关键词: 瘦素；瘦素受体；HTR8-SVneo细胞；分化抑制因子2；增殖

Abstract:

Objective To investigate the effects of leptin on proliferation of HTR8-SVneo cells in vitro and the level of ID(inhibitor of DNA binding and differentiation), and to explore the regulatory mechanism of leptin underlying HTR8-SVneo cell proliferation. Methods The proliferation of HTR8-SVneo cells cultured in vitro under diverse concentrations (0, 10, 50, 100, 250 and 500ng/mL) of leptin was tested by means of MTT. Then, HTR8-SVneo cells were treated with leptin (0, 100 and 500ng/mL), and leptin receptor(leptin R) profiling, ID mRNA profiling and the level of the ID2 protein were detected by RT-PCR, real-time PCR and Western blot, respectively. To observe the effect of leptin on HTR8-SVneo cell proliferation after ID2-siRNA treatment, HTR8-SVneo cells were divided into 6 groups: leptin(0ng/mL), leptin(100ng/mL), leptin(500ng/mL), leptin(0ng/mL)+ID2-siRNA, leptin(100ng/mL)+ID2-siRNA and leptin(500ng/mL)+ID2-siRNA. Results Proliferation of HTR8-SVneo cells was enhanced by high concentrations of leptin (100, 250 and 500ng/mL) compared with leptin (0ng/mL) (P<0.05). Leptin receptor subtypes except HuB219.2 were detected on HTR8-SVneo cells. The up-regulation of ID2 or OB-RL was induced by leptin in a dosedependent manner. ID2-siRNA inhibited expression of the ID2 protein and reversed the enhancement of HTR8-SVneo cell proliferation induced by leptin. Conclusion Leptin enhances HTR8-SVneo cell proliferation by the pathway of leptin R-ID2, which offers new basal data for elucidating the regulatory mechanism of proliferation of trophocytes.

Key words: Leptin; Leptin receptor; HTR8-SVneo cells; Inhibitor of differentiation 2; Proliferation

中图分类号:

R714.12

程欢欢1，王华阳1，董召刚2，徐小飞3，孔北华3，曲迅1. 瘦素对HTR8-SVneo细胞增殖的影响及调控机制[J]. 山东大学学报(医学版), 2012, 50(1): 51-56.

CHENG Huan-huan1, WANG Hua-yang1, DONG Zhao-gang2, XU Xiao-fei3, KONG Bei-hua3, QU Xun1. Effect of leptin on HTR8-SVneo cell proliferation in vitro and its mechanism[J]. JOURNAL OF SHANDONG UNIVERSITY (HEALTH SCIENCES), 2012, 50(1): 51-56.

参考文献

多维度评价

Viewed

Full text

Abstract

Cited

Shared

Discussed

[1]	董坤，张向丽，苏雪莲，刘媛. PPARα和IL-1βmRNA在妊娠期子宫肌层的表达及相关性[J]. 山东大学学报(医学版), 2012, 50(9): 105-.
[2]	徐金娥1,2，王珊1，赵跃然1，陈子江1. 蜕膜组织杀伤细胞免疫球蛋白样受体与不明原因早期复发性流产的相关性[J]. 山东大学学报(医学版), 2009, 47(12): 81-85.
[3]	. 不明原因早期复发性自然流产患者蜕膜组织中杀伤细胞免疫球蛋白样受体2DL1的表达[J]. 山东大学学报(医学版), 2009, 47(7): 81-84.