miR-183靶向TBX3调控PTEN通路对乳腺癌生物学行为影响

发布时间：2018-08-07 07:29

【摘要】：目的乳腺癌早期诊断困难,且转移较早。miR-183在多种肿瘤中存在表达异常的情况,可能影响肿瘤的发生发展过程。为解决乳腺癌早期诊断困难的问题,本研究探讨miR-183在乳腺癌组织中的表达情况,以及对乳腺癌细胞侵袭、迁移能力的影响及其相关机制。方法 PCR检测2015-01-12-2016-03-12四川省人民医院收治的80例乳腺癌组织中miR-183的表达情况。miR-183类似物过表达miR-183,荧光素酶报告基因检测miR-183与T盒转录因子3(T-box transcription factor3,TBX3)的相互作用;蛋白质印迹检测miR-183-mimic处理后,乳腺癌细胞TBX3、第10号染色体同源缺失性磷酸酶-张力蛋白基因(phosphatase and tensin homolog deleted on chromosome ten,PTEN)和磷酸化Akt(phosphorylation-Akt,p-Akt)蛋白的表达情况;Transwell侵袭实验检测miR-183对乳腺癌细胞侵袭能力的影响;划痕实验检测miR-183对乳腺癌细胞迁移能力的影响;裸鼠皮下成瘤检测miR-183对乳腺癌成瘤大小以及体积的影响。结果乳腺癌组织中miR-183表达明显降低,且miR-183的表达水平随乳腺癌病理分期的增加而降低[(85.63±9.28)和(17.56±4.18)],P=0.017;随分化程度的降低,miR-183的表达水平逐渐降低;miR-183的表达随着腋窝淋巴结受累以及受累数目的增多而降低。TBX3是miR-183的直接靶点[(82.24±6.25)和(16.28±6.61)],P=0.028。miR-183可以下调TBX3[(81.4±6.8和19.7±2.1)]和p-AKT[(88.7±7.3)和(24.1±3.1)]蛋白的表达,P=0.008;可以上调PTEN[(62.6±5.3)和(15.6±3.6)]蛋白的表达,P=0.018。miR-183可以抑制乳腺癌细胞的侵袭和迁移能力[(301.5±25.2)和(92.2±12.3)],P=0.005。与NC组相比,miR-183-mimic组荷瘤小鼠肿瘤体积[(2.72±0.13)cm3和(0.35±0.02)cm3]和[(2.55±0.15)g和(0.29±0.03)g]重量都明显变小,P=0.048。结论 miR-183在乳腺癌中起抑癌基因作用,可以下调TBX3的表达,通过PTEN通路抑制乳腺癌细胞侵袭和迁移能力。
[Abstract]:Objective it is difficult to diagnose early breast cancer, and the expression of .miR-183 is abnormal in many kinds of tumors, which may affect the occurrence and development of breast cancer. In order to solve the difficult problem of early diagnosis of breast cancer, this study investigated the expression of miR-183 in breast cancer tissues, the influence of miR-183 on the invasion and migration ability of breast cancer cells and its related mechanism. Methods PCR was used to detect the expression of miR-183 in 80 patients with breast cancer treated in Sichuan Provincial people's Hospital from January to December 2015-06-12. MiR-183 analogue was over-expressed miR-183.The interaction between miR-183 and T-box transcription factor3TBX3 was detected by luciferase reporter gene. Western blot analysis showed that after miR-183-mimic treatment, Expression of TBX3, (phosphatase and tensin homolog deleted on chromosome tenon (PTEN) and phosphorylation-Aktnp-Akt (phosphorylation-Aktnp-Akt) protein in human breast cancer cell line TBX3; Transwell invasion assay was used to detect the effect of miR-183 on the invasive ability of breast cancer cells. The effect of miR-183 on the migration of breast cancer cells was detected by scratch test, and the effect of miR-183 on the size and volume of breast cancer was detected by subcutaneous tumorigenesis in nude mice. Results the expression of miR-183 was significantly decreased in breast cancer. The expression level of miR-183 decreased with the increase of pathological stage of breast cancer [(85.63 卤9.28) and (17.56 卤4.18)], and with the decrease of differentiation degree, the expression of miR-183 decreased gradually with the increase of axillary lymph node involvement and the number of involvement. TBX3 was miR-183. The direct target [(82.24 卤6.25) and (16.28 卤6.61)] P0.028.miR-183 down-regulated the expression of TBX3 [(81.4 卤6.8,19.7 卤2.1)] and p-AKT [(88.7 卤7.3) and (24.1 卤3.1)] protein, and up-regulated the expression of PTEN [(62.6 卤5.3) and (15.6 卤3.6)] protein in breast cancer cells [(301.5 卤25.2) and (92.2 卤12.3)]. Compared with NC group, tumor volume [(2.72 卤0.13) cm3 and (0.35 卤0.02) cm3] and [(2.55 卤0.15) g and (0.29 卤0.03) g] of tumor-bearing mice in miR-183-mimic group were significantly smaller than those in NC group. Conclusion miR-183 acts as a tumor suppressor gene in breast cancer and can down-regulate the expression of TBX3 and inhibit the invasion and migration of breast cancer cells through PTEN pathway.
【作者单位】：四川省人民医院城东病区甲状腺乳腺外科;
【分类号】：R737.9

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