摘要
目的:检测miR-155-5p在不同宫颈疾病患者血清中的表达差异,并分析其对宫颈癌细胞增殖、细胞周期和凋亡的影响,探讨miR-155-5p在宫颈癌发生、发展中的可能作用机制。方法:采用SYBR GreenⅠ实时荧光定量PCR法,检测并分析比较miR-155-5p在不同宫颈疾病患者血清中的表达差异。利用miR-155-5p mimic或inhibitor提高或降低宫颈癌细胞中miR-155-5p的表达。CCK-8法和流式细胞术检测宫颈癌细胞的增殖、细胞周期和凋亡。结果:宫颈癌组血清中miR-155-5p的表达高于宫颈炎组和健康对照组(P<0.05),宫颈上皮内瘤样病变组和宫颈癌组血清中miR-155-5p的表达差异无统计学意义(P>0.05)。与空白组、脂质体组和阴性对照组相比,转染100 nmol/L和200 nmol/L miR-155-5p mimic的SiHa细胞中,S期细胞比例升高,凋亡细胞比例降低(P<0.05)。转染100 nmol/L和200 nmol/L miR-155-5p inhibitor的SiHa细胞中,G2/M期细胞比例明显增多(P<0.05)。结论:(1)宫颈癌患者血清中miR-155-5p表达较健康对照人群上调,可能作为宫颈癌早期诊断的肿瘤分子标志物。(2)miR-155-5p对宫颈癌HeLa细胞增殖、细胞周期和凋亡无明显影响。(3)miR-155-5p可促进宫颈癌SiHa细胞进入S期,并抑制SiHa细胞凋亡,提示miR-155-5p可能在宫颈鳞癌发生、发展中起作用。
AIM: To detect the serum level of miR-155-5p in the patients with different cervical diseases, and to analyze its effects on the proliferation, cell cycle and apoptosis of cervical cancer cells. METHODS: SYBR Green I real-time quantitative PCR was used to detect the level of miR-155-5p in the serum of the patients with different cervical diseases, miR-155-5p mimic or inhibitor was used to increase or decrease the expression of miR-155-5p in cervical cancer cells. The proliferation, cell cycle and apoptosis were measured by CCK-8 assay and flow cytometry. RESULTS: The ser- um level of miR-155-5p in cervical cancer group was higher than that in cervicitis group and healthy group. No statistical difference of the serum miR-155-5p level between cervical intraepithelial neoplasia group and cervical cancer group was ob- served. Compared with blank group, liposome group and negative control group, the proportion of S-phase cells increased and apoptotic cells decreased in SiHa ceils transfected with 100 nmol/L and 200 nmol/L miR-155-5p mimic. The propor- tion of G2/M-phase cells increased significantly in SiHa cells transfected with 100 nmol/L and 200 nmol/L miR-155-5p in- hibitor. CONCLUSION: Compared with healthy controls, the serum level of miR-155-5p in the cervical cancer patients increases, and may act as a novel tumor molecular marker for diagnosis of cervical cancer, miR-155-5p has no significant effect on the proliferation, cell cycle and apoptosis of HeLa cell. miR-155-5p may promote SiHa cells to enter S phase and inhibit the apoptosis of SiHa cells.
出处
《中国病理生理杂志》
CAS
CSCD
北大核心
2013年第12期2193-2200,共8页
Chinese Journal of Pathophysiology
基金
广东省科技计划(No.2011B031800315)