miR-196a靶基因HoxB8在宫颈癌细胞中的表达及临床意义的初步探讨

发布时间：2018-06-02 21:15

本文选题：宫颈鳞癌 + miR-196a　；参考：《广西医科大学》2017年硕士论文

【摘要】：目的探讨miR-196a及其靶基因HoxB8 mRNA在不同宫颈癌细胞株中的表达情况,HoxB8基因对宫颈癌细胞凋亡的影响;探讨HoxB8蛋白在宫颈鳞癌组织中的表达及临床意义。方法1.通过实时荧光定量PCR检测宫颈癌Caski、SiHa、C33-A和HeLa细胞系中miR-196a和HoxB8 mRNA的表达情况;通过q RT-PCR检测miR-196a模拟物转染宫颈癌SiHa细胞后对HoxB8表达的影响;通过HoxB8 siRNA瞬时转染宫颈癌SiHa细胞后检测Caspase3/7的活性;2.收集整理广西医科大学附属肿瘤医院妇瘤科2011年2月至2012年12月行宫颈活检术67例HPV16(+)IIb-IIIb期宫颈鳞癌放化疗或放疗病例的临床病理资料,并在病理科标本库找出其石蜡标本。通过免疫组化染色的方法研究HoxB8的表达情况,并进一步分析其与各临床病理因素及预后的关系。用卡方检验或Fisher精确检验进行与临床病理因素间相关性比较,Kaplan-Meier法计算生存率,Log-rank法进行生存曲线比较,Cox回归模型进行多因素分析。结果1.实时荧光定量PCR结果显示,miR-196a在HPV16阳性的Caski和SiHa细胞系中的表达明显低于HPV16阴性的C33-A和HeLa,差异具有统计学意义(P0.001);与之相对应的,HoxB8在HPV16阳性的Caski和SiHa细胞系中的表达则明显高于HPV16阴性的C33-A和HeLa,差异具有统计学意义(P0.001);miR-196a可下调HoxB8的表达;Capase3/7活力检测结果显示,Capase3/7活力在SiHa细胞HoxB8 si RNA转染组明显高于阴性对照组和正常对照组,差异具有统计学意义(P0.05)。2.HoxB8阴性10例,阳性57例(85%)。染色结果与各临床病理因素相关性分析显示HoxB8与年龄、肿瘤最大径、治疗前SCC、分化程度、FIGO分期均无统计学意义。67例患者失访率19.4%,中位总生存时间为48个月,1年、3年、5年总生存率分别为91%、66%和54%。单因素及多因素生存分析显示,肿瘤最大径4cm、单纯放疗是IIb-IIIb宫颈鳞癌患者预后不良的独立危险因素,差异具有统计学意义(P0.05)。结论miR-196a可能通过下游靶基因HoxB8影响宫颈癌细胞凋亡;HoxB8与HPV16(+)IIb-IIIb期宫颈鳞癌患者年龄、肿瘤最大径、治疗前SCC、分化程度、FIGO分期无明显相关性;肿瘤最大径4cm、单纯放疗HPV16(+)IIb-IIIb宫颈鳞癌患者总生存时间缩短,肿瘤最大径、治疗方式是影响OS的独立危险因素;HoxB8与HPV16(+)IIb-IIIb期宫颈鳞癌患者预后无明显相关性。
[Abstract]:Objective to investigate the expression of miR-196a and its target gene HoxB8 mRNA in different cervical cancer cell lines and the effect of HoxB8 gene on apoptosis of cervical cancer cells, and to explore the expression and clinical significance of HoxB8 protein in cervical squamous cell carcinoma. Method 1. The expression of miR-196a and HoxB8 mRNA in the cervical cancer cell lines Caskius SiHaC33-A and HeLa was detected by real-time fluorescence quantitative PCR, and the effect of miR-196a mimics on HoxB8 expression in cervical cancer SiHa cells was detected by Q RT-PCR. The activity of Caspase3/7 was detected by transient transfection of HoxB8 siRNA into cervical cancer SiHa cells. The clinicopathological data of 67 cases of HPV16 (stage IIb-IIIb) cervix squamous cell carcinoma treated with radiotherapy, chemotherapy or radiotherapy from February 2011 to December 2012 were collected and analyzed. The paraffin specimens were found in the specimen bank of pathology department. Immunohistochemical staining was used to study the expression of HoxB8 and its relationship with clinicopathological factors and prognosis. The correlation between clinical and pathological factors was compared by chi-square test or Fisher accurate test. Kaplan-Meier method was used to calculate survival rate and Log-rank method was used to compare survival curve and Cox regression model was used for multivariate analysis. Result 1. The results of real-time fluorescence quantitative PCR showed that the expression of miR-196a in HPV16 positive Caski and SiHa cell lines was significantly lower than that in HPV16 negative C33-A and HeLa cells, the difference was statistically significant (P0.001), and the corresponding expression of HoxB8 in HPV16 positive Caski and SiHa cell lines was significantly lower than that in HPV16 negative Caski and SiHa cell lines. It was significantly higher than that of C33-A and HeLa-negative HPV16, and the difference was statistically significant. P0.001 miR-196a could down-regulate the expression of HoxB8 and Capase3 / 7 activity. The results showed that the activity of capase3 / 7 in SiHa cell HoxB8 si RNA transfection group was significantly higher than that in SiHa cell HoxB8 si RNA transfection group and normal control group. The difference was statistically significant (P 0.05) .2.HoxB8 was negative in 10 cases and positive in 57 cases (85%). The correlation analysis between staining results and clinicopathological factors showed that HoxB8 was associated with age and the maximum diameter of tumor. There was no significant difference in SCC, differentiation and Figo stage before treatment. The median survival time was 48 months, 1 year, 3 years and 5 years. The overall survival rate was 91 66% and 54% respectively. Univariate and multivariate survival analysis showed that the maximum diameter of the tumor was 4 cm and radiotherapy alone was an independent risk factor for poor prognosis in patients with IIb-IIIb cervical squamous cell carcinoma. The difference was statistically significant (P 0.05). Conclusion miR-196a may affect apoptosis of cervical cancer cells by downstream target gene HoxB8. There is no significant correlation between HoxB8 and HPV16 (stage IIb-IIIb) cervical squamous cell carcinoma patients age, tumor maximum diameter, pre-treatment SCC, differentiation degree and Figo stage. The maximum diameter of tumor was 4 cm, the total survival time of patients with cervical squamous cell carcinoma of HPV16 was shortened and the maximum diameter of tumor was shortened by radiotherapy alone. The independent risk factor of affecting OS was HoxB8 and the prognosis of patients with squamous cell carcinoma of cervix of HPV16 was not significantly correlated with that of HPV16 (stage IIb-IIIb).
【学位授予单位】：广西医科大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R737.33

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