| Angiomotin基因沉默增强乳腺癌干细胞特性及其机制探讨 |
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| 引用本文： | 吕美玲,刘培军,杨 瑾.Angiomotin基因沉默增强乳腺癌干细胞特性及其机制探讨[J].现代肿瘤医学,2022,0(9):1560-1566. |
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| 作者姓名： | 吕美玲 刘培军 杨 瑾 |
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| 作者单位： | 1.长安大学医院内科,陕西 西安 710064;
2.西安交通大学第一附属医院肿瘤内科,陕西 西安 710061 |
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| 摘 要： | 目的:研究Angiomotin(Amot)基因沉默后对乳腺癌干细胞特性的影响，初步评估Amot在乳腺癌中异常表达的分子机制。方法:运用免疫组化、Western blot及RT-qPCR技术在乳腺癌组织及乳腺癌细胞株中验证Amot的表达，进而运用RNA干扰技术特异性地阻断Amot在乳腺癌高表达细胞株MCF-7中的表达，以期了解Amot沉默后对乳腺癌干细胞特性的影响。结果:114例乳腺癌组织中，97例阳性表达，阳性率达85.09%；92例非癌组织中，阳性表达10例，阳性率为10.87%，两组差异具有显著统计学意义（P＜0.001）。Amot在乳腺癌组织中高表达，定位在细胞核和胞浆中，主要定位于细胞核；组织芯片癌旁组织中低表达。MCF-7细胞系经Amot沉默后，细胞形态学上发生上皮间质表型转化。Amot基因沉默后，乳腺癌MCF-7干细胞相关分子表型标志CD24/CD44双标阳性率的变化:CON组:CD24（11.1±0.55）%、CD44<（0.1±0.48）%；NC组:CD24（12.4±0.62）%、CD44<（0.08±0.7）%；KD组:CD24<（0.1±0.08）%、CD44（81.0±2.02）%。乳腺癌干细胞相关分子表型标志物CD24表达降低，CD44表达明显增强，差异有统计学意义（P＜0.001）。肿瘤球形成率:CON组2.6%，NC组2.8%，KD组9.75%；乳腺癌细胞在Amot沉默后形成“肿瘤球”能力增强，差异具有统计学意义（P＜0.01）。干细胞特性相关基因C-myc、Sox-2表达增加；上皮蛋白E-cadherin表达减弱，间皮蛋白N-cadherin、Vimentin、a-SMA、Snail表达明显增加，差异均有统计学意义（P＜0.05）。同时发现，MCF-7细胞系经Amot 沉默后，Hippo-YAP通路中YAP及YAP/TAZ表达降低，YAP上游LATS1表达降低，差异有统计学意义（P＜0.01）。结论:Amot基因沉默增强乳腺癌干细胞特性，诱发乳腺癌细胞发生EMT。
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| 关 键 词： | 乳腺癌 MCF-7细胞 Angiomotin 肿瘤干细胞 EMT Hippo |
Angiomotin gene silencing enhances the characteristics of breast cancer stem cells and its mechanism |
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| LYU Meiling,LIU Peijun,YANG Jin.Angiomotin gene silencing enhances the characteristics of breast cancer stem cells and its mechanism[J].Journal of Modern Oncology,2022,0(9):1560-1566. |
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| Authors: | LYU Meiling LIU Peijun YANG Jin |
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| Affiliation: | 1.Department of Internal,Chang'an University Hospital,Shaanxi Xi'an 710064,China;2.Department of Oncology,First Affiliated Hospital of Xi'an Jiaotong University,Shaanxi Xi'an 710061,China. |
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| Abstract: | Objective:To investigate the effect of Angiomotin(Amot) silencing on the characteristics of breast cancer stem cells,and to preliminarily evaluate the molecular mechanism of Amot abnormal expression in breast cancer.Methods:Immunohistochemistry,Western blot and RT-qPCR were used to verify the expression of Amot in breast cancer tissues and breast cancer cell lines.Then shRNA was applied to block and silence the expression of Amot in the breast cancer cell lines,and the effects of the silenced Amot on stem cell properties of breast cancer cell lines were studied.Results:Among 114 cases of breast cancer,97 cases were positively expressed,with a positive rate of 85.09%.Among the 92 non-cancerous tissues,positive expression was found in 10 cases,with a positive rate of 10.87%,and the difference was statistically significant(P＜0.001).Amot is highly expressed in breast cancer tissues and is located in the nucleus and cytoplasm,mainly in the nucleus.While low expression in paracancer tissues.After Amot silencing,MCF-7 cell lines underwent morphological epithelial mesenchymal phenotypic transformation.After Amot silencing,the molecular phenotypic marker of breast cancer stem cells showed low expression of CD24 and significantly increased expression of CD44(P＜0.001).The tumor spheres formation rate was CON 2.6%,NC 2.8% and KD 9.75% respectively.The ability of breast cancer cells to form "tumor spheres" was enhanced after Amot silencing(P＜0.01).The expression of C-myc and Sox-2,factors related to the characteristics of stem cells,were increased.The expression of E-cadherin was decreased,while N-cadherin,Vimentin,a-SMA and Snail were significantly increased,and the differences were statistically significant(P＜0.05).It was also found that after Amot silencing in MCF-7 cell lines,the expression of YAP and YAP/TAZ in Hippo-YAP pathway was decreased,and the expression of LATS1 was decreased,with statistically significant difference(P＜0.01).Conclusion:Amot gene silencing enhances the characteristics of breast cancer stem cells and induces the EMT of breast cancer cells. |
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| Keywords: | breast cancer MCF-7 cell Angiomotin CSCs EMT Hippo |
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