摘要
目的探讨肿瘤干细胞在乳腺癌患者腋窝肿大淋巴结中微转移灶及微小RNA30a(miR30a)对其侵袭能力的影响,初步探讨miRNAs抗乳腺癌治疗的可行性。方法从乳腺癌患者腋窝肿大淋巴结中分离、培养肿瘤干细胞样乳腺癌细胞。合成miR30a寡核苷酸片段,应用腺病毒将该片段转染人原代乳腺癌细胞,同时设乳腺癌MDA-MB-231细胞株为试验对照,每组细胞均设空载体组、空白对照组,以荧光显微镜评估转染效率。以Transwell小室体外侵袭试验检测转染前后肿瘤细胞增殖和侵袭力的变化。以Western blot分别检测ALDH1、Vimentin和N-Cadherin蛋白表达。结果 Transwell小室体外侵袭试验显示空白对照组中原代乳腺癌细胞较MDA-MB-231细胞株侵袭力强,其侵袭指数分别为(75.3±3.2)%,(58.4±2.8)%,两者差异有统计学意义(P<0.05),而转染miR30a后这两种细胞体外侵袭力均明显减弱,其侵袭指数分别为(21.4±1.9)%,(28.2±2.3)%,与各自空白对照组相比差异有统计学意义(P<0.05)。结论乳腺癌患者部分腋窝增生肿大的淋巴结中ALDH表达增高,可能存在肿瘤微转移,术中应完全清扫为宜。miR30a抑制了肿瘤干性基因表达和侵袭能力。
Objective To investigate the micrometastatic lesion of tumor stem cells in the axillary swollen lymph nodes of breast cancer patients and the influence of miR30 aon its invasive ability,and to explore the feasibility of miRNAs anti-breast cancer treatment.Methods The tumor stem cell-like breast cancer cells were separated from the axillary swollen lymph nodes in breast cancer patients and cultured.miR30 aoligonucleotide fragment was synthesized and transfected into human primary generation breast cancer cells by using adenovirus,meanwhile the breast cancer cell line MDA-MB-231 was taken as the experimental control and the transfection efficiency was assessed by the fluorescence microscopy.The changes of tumor cell proliferation and invasiveness before and after transfection were detected by the Transwell chamber in vitro invasion assay.Western blot was used to detect the ALDH1,Vimentin and N-Cadherin protein expression.Results The Transwell chamber in vitro invasion assay showed that the primary generation breast cancer cell had more strong invasive ability than MDA-MB-231 cell line,their invasion indexes were（75.3±3.2）% and（58.4±2.8）%respectively,the difference was statistically significant（P〈0.05）,and after transfecting miR30 a,the in vitro invasiveness ability in these two kinds of cells were significantly weakened and their invasion indexes were（21.4±1.9）%and（28.2±2.3）% respectively,the difference compared with the control group showed the statistical significance（P〈0.05）.Conclusion The ALDH expression in partial axillary hyperplasia and swollen lymph nodes in the patients with breast cancer is increased,and the tumor micrometastasis may exist,which should be completely cleaned in operation.miR30 ainhibits the expression and invasive ability of tumor stem gene.
出处
《重庆医学》
CAS
北大核心
2016年第12期1605-1607,1611,共4页
Chongqing medicine
基金
重庆市科技计划项目科研基金支持(CSTC2011JJA10038)
