MicroRNA-96在宫颈癌组织中的表达及其与化疗敏感性关系的研究

发布时间：2018-04-26 01:25

本文选题：小分子RNA-96 + 宫颈癌　；参考：《广州中医药大学》2014年硕士论文

【摘要】：目的：探讨小分子RNA-96(microRNA-96, miR-96)在子宫颈癌(Uterine cervical cancer, UCC)组织中的表达情况及其与临床病理特征之间的关系；探讨新辅助化疗对宫颈鳞状细胞癌组织中miR-96表达的影响,及miR-96的表达变化情况与新辅助化疗疗效、敏感性、临床常见的病理特征之间的关系,以评估其在化疗疗效及敏感性方面的预测价值；探讨宫颈癌患者组织中miR-96的表达水平与宫颈癌中医辨证分型之间的关系,评价中西医结合治疗方法应用于宫颈癌临床治疗中的价值和前景。方法：采用茎环实时荧光定量聚合酶链反应技术(stem-loop quantitative real timePCR,qPCR)测定52例子宫颈恶性肿瘤患者的肿瘤组织、28例宫颈正常患者的宫颈组织中miR-96的表达；同法,检测28例(Ⅰb2期-Ⅱa期)宫颈鳞状细胞癌患者新辅助化疗前、后肿瘤组织中miR-96的表达；统计分析miR-96的表达水平与临床病理特征之间的关系及miR-96的表达变化与新辅助化疗的疗效、敏感性、临床病理特征之间的相关性；运用中医辨证分型对宫颈癌组织中miR-96的表达情况进行分析。结果： 1.52例宫颈癌患者肿瘤组织中miR-96的相对表达量为45.59(15.19,139.61)显著高于28例正常宫颈组织0.94(0.78,1.11),差异具有统计学意义(p=0.000)。 2.MiR-96在宫颈癌患者组织中的相对表达量与临床病理特征(临床分期,组织学类型,细胞学分级,浸润深度,淋巴结转移)关系密切：其在中、低分化癌中的相对表达量61.61(17.74,183.42)显著高于高分化癌16.78(6.13,66.67),差异有统计学的意义(p=0.005)；在腺癌中相对表达量为155.67(57.77,270.61),明显高于鳞癌27.28(13.35,91.00),(p=0.004)；临床Ⅰ期、Ⅱ期、Ⅲ～Ⅳ期子宫颈癌患者组织中miR-96的相对表达量分别为15.41(7.52,26.26)、85.15(68.32,194.76)462.08(422.57,1028.65),Ⅲ～Ⅳ期显著高于Ⅰ期、Ⅱ期(p=0.000、p=0.001),Ⅱ期显著高于Ⅰ期,且差异均具有统计学的意义(P=0.000)；淋巴结有转移组的相对表达量为163.46(28.95,205.62)明显高于无淋巴结转移组14.16(6.99,22.61),(p=0.000)；浸润深度：≥1/2间质的相对表达量为24.72(14.44,141.79),显著高于1/2间质者13.79(5.14,25.649),(p=0.025)；而与患者年龄无明显关系(p=0.385)。 3.28例宫颈鳞状细胞癌患者新辅助化疗后miR-96的表达水平明显下降；新辅助化疗前宫颈癌肿瘤组织中miR-96的相对表达量是化疗后肿瘤组织中的3.40(2.01,6.86)倍,差异具有统计学的意义(p=0.000)。 4.宫颈鳞状细胞癌患者新辅助化疗前、后肿瘤组织中miR-96表达变化的相对定量结果与临床病理特征之间关系密切：细胞分化程度为中、低分化者2.51(1.21,33.26)低于高分化者8.22(5.21,15.39)(p=0.000)；间质浸润深度1/2间质者5.66(4.06,14.09)明显高于浸润深度≥1/2间质者1.91(0.90,2.96)(p=0.000)；有淋巴结转移者1.79(0.86,2.64)明显低于无淋巴转移者5.43(4.04,12.84)(p=0.000)；而年龄≤45岁、45岁,临床分期为Ⅰb2期、Ⅱa期患者,化疗前后miR-96表达变化均没有统计学差异(p0.05)。 5.新辅助化疗有效组(CR+PR)同无效组(SD+PD)相比较：有效组化疗后肿瘤组织中的miR-96表达水平明显下降(p=0.000),无效组化疗前、后肿瘤组织中miR-96的表达水平变化无统计学差异(p0.05)；新辅助化疗前有效组肿瘤组织中miR-96的表达水平明显低于无效组(p=0.001),新辅助化疗后有效组肿瘤组织中miR-96的表达水平也明显低于无效组(p=0.001)。 6.宫颈鳞状细胞癌患者行新辅助化疗前肿瘤组织中miR-96的相对表达量与三维超声参数血流指数(FI)、血管形成指数(VI)、血管血流指数(VFI)、化疗后残余肿瘤体积百分比呈明显正相关(p0.05),而与化疗前肿瘤体积无明显相关性(p0.05)。 7.宫颈癌患者组织中miR-96的表达水平与中医辨证分型之间的关系：肝肾阴虚、脾肾阳虚型患者肿瘤组织中miR-96的表达水平明显高于肝郁气滞型、湿热瘀毒型,差异有统计学的意义(p0.05)；而脾肾阳虚型与肝肾阴虚型患者肿瘤组织中miR-96的表达,肝郁气滞型与湿热瘀毒型患者肿瘤织中miR-96的表达均无统计学差异(p0.05)。结论： 1.MiR-96在宫颈癌组织中异常高表达,其表达水平与患者临床病理特征之间关系密切,提示其可能在宫颈癌的发生发展中发挥致癌基因的作用,并且与宫颈癌的演进及不良预后密切相关。 2.新辅助化疗能有效降低miR-96在子宫颈恶性肿瘤组织中的表达,化疗前肿瘤组织中miR-96的表达水平与化疗的敏感性、临床疗效关系密切,因此测定化疗前宫颈癌组织中miR-96的表达水平对新辅助化疗疗效及敏感性预测具有一定参考价值。 3.宫颈癌患者肿瘤组织中miR-96的表达水平与中医辨证分型密切相关：脾肾阳虚型、肝肾阴虚证型中的表达水平明显高于肝郁气滞型、湿热瘀毒证型。
[Abstract]:Objective:
To investigate the expression of small molecule RNA-96 (microRNA-96, miR-96) in the tissues of Uterine cervical cancer (UCC) and its relationship with the clinicopathological features, and to explore the effect of neoadjuvant chemotherapy on the expression of miR-96 in cervical squamous cell carcinoma, and the changes in the expression of miR-96 and the efficacy and sensitivity of the neoadjuvant chemotherapy. The relationship between the common pathological features of the clinic to evaluate the predictive value of the therapeutic effect and sensitivity in chemotherapy, the relationship between the expression of miR-96 in the tissues of cervical cancer patients and the TCM syndrome differentiation of cervical cancer, and to evaluate the value and Prospect of the combination of traditional Chinese and Western medicine in the clinical treatment of cervical cancer.
Stem-loop quantitative real timePCR (qPCR) was used to determine the expression of miR-96 in 52 cases of cervical malignant tumor, 28 cases of cervical normal patients and 28 cases of cervical squamous cell carcinoma (I B2 phase II a phase) before and after neoadjuvant chemotherapy and posterior swelling in 28 cases of cervical squamous cell carcinoma. The expression of miR-96 in the tumor tissue; statistical analysis of the relationship between the expression level of miR-96 and the clinicopathological features and the correlation between the expression of miR-96 and the curative effect, sensitivity and clinicopathological features of the neoadjuvant chemotherapy, and the analysis of the expression of miR-96 in cervical cancer by TCM syndrome differentiation.
The relative expression of miR-96 in the tumor tissues of 1.52 patients with cervical cancer was 45.59 (15.19139.61) significantly higher than that in 28 cases of normal cervical tissue (0.78,1.11), and the difference was statistically significant (p=0.000).
The relative expression of 2.MiR-96 in the tissues of patients with cervical cancer was closely related to the clinicopathological features (clinical stage, histological type, cytological classification, infiltration depth, lymph node metastasis). In the middle, the relative expression of 61.61 (17.74183.42) in the low differentiated carcinoma was significantly higher than that of the highly differentiated carcinoma (6.13,66.67), and the difference was statistically significant (p=0 The relative expression of.005 in adenocarcinoma was 155.67 (57.77270.61), which was significantly higher than that of squamous carcinoma (13.35,91.00), (p=0.004). The relative expression of miR-96 in the tissues of patients with stage I, stage II, and stage III to IV of cervical cancer was 15.41 (7.52,26.26), 85.15 (68.32194.76) 462.08 (422.571028.65) and stage III to IV significantly higher than stage I, stage II (P). =0.000, p=0.001), the stage II was significantly higher than phase I, and the difference was statistically significant (P=0.000), the relative expression of the lymph node metastasis group was 163.46 (28.95205.62) significantly higher than that in the non lymph node group (6.99,22.61), (p=0.000), and the infiltration depth was 24.72 (14.44141.79), which was significantly higher than that of 1/2 (14.44141.79). The interstitial mass was 13.79 (5.14,25.649), (p=0.025), but not significantly related to the age of the patients (p=0.385).
The expression level of miR-96 in 3.28 cases of cervical squamous cell carcinoma was significantly decreased after neoadjuvant chemotherapy, and the relative expression of miR-96 in cervical cancer tissues before neoadjuvant chemotherapy was 3.40 (2.01,6.86) times of the tumor tissue after chemotherapy, and the difference was statistically significant (p=0.000).
4. before the neoadjuvant chemotherapy of cervical squamous cell carcinoma, the relative quantitative results of miR-96 expression in the posterior tumor tissues were closely related to the clinicopathological features: the degree of cell differentiation was in the middle, the low differentiation 2.51 (1.21,33.26) was 8.22 (5.21,15.39) lower than that of the high differentiation (p= 0), and the interstitial infiltrating depth of the interstitial 1/2 was 5.66 (4.06,14.09). 1.91 (0.90,2.96) (p=0.000) and 1.79 (0.86,2.64) with lymph node metastases were significantly lower than those of 1/2 (0.86,2.64) with lymph node metastases, and 5.43 (4.04,12.84) (p=0.000) were lower than those without lymphatic metastasis, while the age was less than 45 years and 45 years old, and the clinical stage was I B2 stage, and there was no statistical difference in the changes of miR-96 expression in the patients with stage II a (P0.05).
5. the effective group of neoadjuvant chemotherapy (CR+PR) was compared with the ineffective group (SD+PD): the expression level of miR-96 in the tumor tissues of the effective group decreased significantly (p=0.000). There was no significant difference in the expression of miR-96 in the tumor tissues (P0.05) before chemotherapy (P0.05); the expression level of miR-96 in the tumor tissues before the neoadjuvant chemotherapy was clear. The expression level of miR-96 in the effective group after neoadjuvant chemotherapy was significantly lower than that in the ineffective group (p=0.001), which was significantly lower than that in the ineffective group (p=0.001).
6. the relative expression of miR-96 in the tumor tissue of the cervical squamous cell carcinoma patients before the neoadjuvant chemotherapy and the three dimensional ultrasound parameter blood index (FI), the angiogenic index (VI), the blood vessel blood flow index (VFI), the percentage of the residual tumor volume after chemotherapy (P0.05), and no significant correlation with the tumor volume before chemotherapy (P0.05).
7. the relationship between the expression level of miR-96 in the tissues of the patients with cervical cancer and the syndrome differentiation of traditional Chinese medicine: the expression level of miR-96 in the tumor tissues of the patients with spleen and kidney yang deficiency is significantly higher than that of the liver qi stagnation, and the difference is statistically significant (P0.05), while the spleen kidney yang deficiency and the liver kidney yin deficiency patients are in the tumor tissue of miR-9. 6, there was no significant difference in the expression of miR-96 in tumor tissue between liver qi stagnation type and damp heat stasis type (P0.05).
1.MiR-96 is highly expressed in cervical cancer, and its expression level is closely related to the clinicopathological features of the patients, suggesting that it may play the role of the oncogene in the development of cervical cancer, and is closely related to the evolution and poor prognosis of cervical cancer.
2. neoadjuvant chemotherapy can effectively reduce the expression of miR-96 in the malignant tumor tissues of the cervix. The expression of miR-96 in the tumor tissue before chemotherapy is closely related to the sensitivity of chemotherapy and the clinical efficacy. Therefore, the determination of the expression level of miR-96 in the cervical cancer tissues before chemotherapy is of reference value for the therapeutic effect and sensitivity prediction of the neoadjuvant therapy.
3. the expression level of miR-96 in the tumor tissues of the patients with cervical cancer is closely related to the TCM syndrome differentiation: the expression level in the spleen kidney yang deficiency type, the liver kidney yin deficiency syndrome type is obviously higher than the liver qi stagnation type, the damp heat stasis syndrome type.

【学位授予单位】：广州中医药大学
【学位级别】：硕士
【学位授予年份】：2014
【分类号】：R737.33

【参考文献】