摘要
目的:体外研究雌激素膜受体GPR30对宫颈癌细胞生长的影响及其作用机制。方法:选择宫颈腺癌HeLa和宫颈鳞癌SiHa细胞株,分别用GPR30特异性激动剂G1和拮抗剂G15处理宫颈癌细胞株。RT-PCR、Western blot法检测处理前后宫颈癌HeLa与SiHa细胞中GPR30、TLR3 mRNA及其蛋白表达变化;MTT法检测G1、G15及Poly I:C处理对宫颈癌细胞生长的影响。结果:(1)HeLa细胞中GPR30表达量高于SiHa细胞。G1处理后HeLa、SiHa细胞中GPR30 mRNA及其蛋白表达水平增高,与对照组比较,差异有统计学意义(P<0.05;P<0.05);G15处理后,HeLa、SiHa细胞中GPR30 mRNA及其蛋白表达水平降低,与对照组比较,差异有统计学意义(P<0.05;P<0.05)。(2)HeLa、SiHa细胞中TLR3 mRNA表达量分别为(0.5327±0.05373)、(0.3526±0.05774),蛋白表达量分别为(0.3572±0.097039)、(0.5002±0.09718)。G1能降低He La、Si Ha细胞中TLR3mRNA及其蛋白表达水平,G1 10-6mol/L处理组与对照组比较差异有统计学意义(P均<0.05)。G15能增高HeLa、SiHa细胞中TLR3 mRNA及其蛋白表达水平,G15 10-5mol/L处理组与对照组比较,差异有统计学意义(P均<0.05),与Poly I:C处理组比较差异无统计学意义(P>0.05)。(3)10-6mmol/L、10-5mmol/L G1分别处理后,宫颈癌HeLa、SiHa细胞生长增殖率分别为(16.68±5.86)%、(26.67±3.25)%及(14.99±6.43)%、(22.72±1.77)%,与空白对照组相比,差异有统计学意义(P均<0.05)。10-6mmol/L、10-5mmol/L G15分别处理后,宫颈癌HeLa、SiHa细胞的生长抑制率分别为(21.09±2.32)%、(22.99±3.15)%及(15.86±6.49)%、(19.18±2.61)%,与空白对照组相比,差异有统计学意义(P均<0.05)。结论:宫颈癌细胞中存在雌激素膜受体GPR30表达,宫颈腺癌细胞中GPR30表达量高于宫颈鳞癌细胞,体外调节GPR30表达可影响宫颈癌细胞生长。抑制GPR30表达可通过上调TLR3表达而抑制宫颈癌细胞生长,GPR30可能成为宫颈癌治疗的新靶点。
Objective: To investigate the effect and mechanism of estrogen receptor GPR30 on the growth of cervical cancer cells in vitro. Methods: Cevrical adenocarcinoma cell line Hela and cervical squamous cancer cell line Siha were treated respectively by GPR30 specific agonist G1 and antagonist G15. The expressions of GPR30 and TLR3 mRNA and their protein levels were detected by RT-PCR and Western-blot. The effect on cell growth was measured by MTT. Result:(1) The expression of GPR30 was higher in Hela than that of Siha. The mRNA and protein expressions of both Hela and Siha increased after treated by G1( P〈0. 05; P〈0.05) and decreased by G15( P〈0. 05,P〈0. 05).( 2) The mRNA expressions of GPR30 in Hela and Siha were( 0. 5327 ± 0. 05373) and( 0. 3526 ± 0. 05774). And the protein expressions were( 0. 3572 ± 0. 097039) and( 0. 5002 ± 0. 09718). G1 can reduce HeLa and SiHa cells TLR3 mRNA and protein expression levels. Compared with the control group,the difference was statistically significant after G1 10^-6mol / L treatment( P〈0. 05). While G15 can increase HeLa and SiHa cells TLR3 mRNA and protein expression levels. Compared with the control group,the difference was statistically significant after G1 10^-5mol / L treatment( P〈0. 05). Compared with the group treated by Poly I: C,the difference was not statistically significant( P〈0. 05).( 3) After treated by G1 with a concentration of 10^-6mmol / L and 10^-5mmol / L,the polification rates of Hela and Siha were( 16. 68 ± 5. 86) %,( 26. 67 ± 3. 25) % and( 14. 99 ± 6. 43) %,( 22. 72 ± 1. 77) %. Compared with the control group,the difference was statistically significant( P〈0. 05). Treated by G15 of 10^-6mmol / L and 10^-5mmol / L,the inhibition rates were( 21.09 ± 2. 32) %,( 22. 99 ± 3. 15) % and( 15. 86 ± 6. 49) %,( 19. 18 ± 2. 61) %. Compared with the control group,the difference was statistically significant( P〈0. 05; P〈0. 05; P〈0. 05; P〈0. 05). Conclusion: GPR30 expressed in cervical cancer cells. The expression levels in cervical adenocarcinoma were higher than those in cervical squamous cell carcinoma. Regulating GPR30 expressions in vitro could influence the growth of cervical cancer cells. Inhibition of GPR30 expressions could inhibit the growth of cervical cancer cells by up-regulating TLR3 expression.GPR30 may become a new target for cervical cancer treatment.
出处
《现代妇产科进展》
CSCD
北大核心
2015年第1期18-21,共4页
Progress in Obstetrics and Gynecology