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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 study the effect of estrogen receptor GPR30 on the growth of cervical carcinoma cells in vitro and its mechanism. Methods: Cervical adenocarcinoma HeLa and cervical squamous cell carcinoma SiHa were selected and treated with GPR30-specific agonist G1 and antagonist G15 respectively. The mRNA and protein expressions of GPR30 and TLR3 in cervical cancer HeLa and SiHa cells were detected by RT-PCR and Western blot. The effects of G1, G15 and Poly I: C on the growth of cervical cancer cells were detected by MTT assay. Results: (1) The expression of GPR30 in HeLa cells was higher than that in SiHa cells. The expression of GPR30 mRNA and protein in HeLa and SiHa cells after G1 treatment was significantly higher than that in control group (P <0.05; P <0.05). After G15 treatment, the expression of GPR30 mRNA and its protein Compared with the control group, the difference was statistically significant (P <0.05; P <0.05). (2) The expression levels of TLR3 mRNA in HeLa and SiHa cells were (0.5327 ± 0.05373) and (0.3526 ± 0.05774), respectively, and the protein expression levels were (0.3572 ± 0.097039) and (0.5002 ± 0.09718), respectively. G1 reduced the expression of TLR3mRNA and protein in HeLa and SiHa cells. The difference was statistically significant between G110-6mol / L group and control group (all P <0.05). G15 could increase the expression of TLR3 mRNA and protein in HeLa and SiHa cells. There was significant difference between G15 10-5mol / L treatment group and control group (P <0.05), and the difference was significant compared with Poly I: C treatment group No statistical significance (P> 0.05). (3) After treated with 10-6mmol / L and 10-5mmol / L G1 respectively, the proliferation rates of cervical cancer HeLa and SiHa were (16.68 ± 5.86)%, (26.67 ± 3.25)% and (14.99 ± 6.43)%, respectively , (22.72 ± 1.77)%, respectively. Compared with the blank control group, the difference was statistically significant (all P <0.05). The growth inhibition rates of cervical cancer HeLa and SiHa cells were (21.09 ± 2.32)%, (22.99 ± 3.15)% and (15.86 ± 6.49)%, respectively after treatment with 10-6mmol / L and 10-5mmol / 19.18 ± 2.61)%, respectively. Compared with the blank control group, the difference was statistically significant (all P <0.05). Conclusion: The expression of estrogen receptor GPR30 in cervical cancer cells is higher than that in cervical squamous carcinoma cells, and the expression of GPR30 in cervical adenocarcinoma cells can affect the growth of cervical cancer cells. Inhibition of GPR30 expression can inhibit the growth of cervical cancer cells by up-regulating the expression of TLR3, and GPR30 may be a new therapeutic target for cervical cancer.