| 韩 拓,肖 栋,陈 亮.ROR1-AS1调控Notch1信号通路对乳腺癌MCF-7细胞增殖和凋亡影响[J].肿瘤学杂志,2022,28(1):40-45. |
| ROR1-AS1调控Notch1信号通路对乳腺癌MCF-7细胞增殖和凋亡影响 |
| Effect of ROR1-AS1 Regulating Notch1 Signaling Pathway on Proliferation and Apoptosis of Breast Cancer MCF-7 Cells |
| 投稿时间：2021-03-01 |
| DOI：10.11735/j.issn.1671-170X.2022.01.B008 |
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| 中文关键词: ROR1-AS1 乳腺癌 上皮-间质转化 凋亡 Notch1信号通路 |
| 英文关键词:ROR1-AS1 breast cancer epithelial-mesenchymal transition apoptosis Notch1 signaling pathway |
| 基金项目:陕西省自然科学基础研究计划［一般项目（面上）(2020JM?鄄393)］ |
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| 中文摘要: |
| 摘 要：［目的］ 研究酪氨酸蛋白激酶跨膜受体 1 反义 RNA 1（tyrosine protein kinase transmembrane receptor 1 antisense RNA 1，ROR1-AS1）调控Notch1信号通路对乳腺癌MCF-7细胞增殖、凋亡及上皮-间质转化(epithelial-mesenchymal transition，EMT)的影响。［方法］ 实时荧光定量PCR方法检测乳腺癌MCF-7、MDA-MB-231、MDA-MB-361细胞和正常乳腺上皮MCF10A细胞中ROR1-AS1表达变化。乳腺癌MCF-7细胞分成对照、sh-NC（转染shRNA 对照）、sh-ROR1-AS1（转染ROR1-AS1 shRNA）、sh-ROR1-AS1+Jagged1组（转染ROR1-AS1 shRNA，Notch1通路激活剂Jagged1处理）。CCK-8实验检测细胞增殖，流式细胞术检测细胞凋亡，Transwell小室检测细胞迁移和侵袭，Western blot方法检测细胞中活化的半胱氨酰天冬氨酸特异性蛋白酶9（cleaved cysteinyl aspartate-specific protease-9，C-Caspase-9）、上皮型钙黏蛋白（E-cadherin）、活化的半胱氨酰天冬氨酸特异性蛋白酶3（cleaved cysteinyl aspartate-specific protease-3，C-Caspase-3）、神经型钙黏蛋白（N-cadherin）、Notch1、Hes1蛋白表达。 ［结果］ 乳腺癌MCF-7、MDA-MB-231、MDA-MB-361细胞中ROR1-AS1水平明显高于正常乳腺上皮MCF10A细胞（P<0.05）。与对照组、sh-NC组比较，sh-ROR1-AS1组乳腺癌MCF-7细胞存活率降低(100.00%±9.82%，99.74%±12.05%，58.91%±6.33%)，细胞凋亡率升高(4.81%±0.26%，4.93%±0.34%，20.58%±2.11%），细胞迁移数目( 225.36±12.84个，224.89±19.56个，140.89±13.64个）和侵袭数目(180.47±16.32个，177.54±16.92个，110.44±10.87个)减少，细胞中C-Caspase-9、E-cadherin、C-Caspase-3蛋白表达水平增加，N-cadherin、Notch1、Hes1蛋白表达水平降低（P<0.05）。与sh-ROR1-AS1组比较，sh-ROR1-AS1+Jagged1组乳腺癌MCF-7细胞存活率升高(100.00%±11.58% vs 147.36%±12.05%），细胞凋亡率降低(19.54%±1.74% vs 8.36%±0.75%)，细胞迁移数目(139.58±12.78个 vs 175.26±16.94个)和侵袭数目（107.45±9.35个 vs 162.04±13.67个）均升高，C-Caspase-9、E-cadherin、C-Caspase-3蛋白水平降低，N-cadherin、Notch1、Hes1蛋白水平升高（P<0.05）。［结论］ 下调ROR1-AS1通过抑制Notch1信号减弱乳腺癌MCF-7细胞增殖、迁移、侵袭和EMT能力，激活细胞凋亡。 |
| 英文摘要: |
| Abstract：［Objective］ To investigate the effect of long non-coding RNA tyrosine protein kinase transmembrane receptor 1 antisense RNA 1（lncRNA ROR1-AS1） regulating Notch1 signaling pathway on the proliferation， apoptosis and epithelial-mesenchymal transition(EMT) of breast cancer cells. ［Methods］ The expression of ROR1-AS1 in human breast cancer MCF-7， MDA-MB-231， MDA-MB-361 cells and normal breast epithelial MCF10A cells were detected by real-time PCR method. Breast cancer MCF-7 cells were transfected with shRNA-NC(sh-NC group)， ROR1-AS1 shRNA(sh-ROR1-AS1 group) or ROR1-AS1 shRNA and Notch1 pathway activator Jagged1(sh-ROR1-AS1+Jagged1 group). Cell proliferation was detected with CCK-8 method， cell apoptosis was detected with flow cytometry， cell migration and invasion was detected with transwell assay， the expression of C-Caspase-9， E-cadherin， C-Caspase-3， N-cadherin， Notch1 and Hes1 proteins was detected with Western blot.［Results］ The expression levels of ROR1-AS1 in breast cancer MCF-7， MDA-MB-231 and MDA-MB-361 cells were significantly higher than that in normal breast epithelial MCF10A cells(P<0.05). Compared with the control and sh-NC groups， in sh-ROR1-AS1 group the cell survival rate decreased(100.00%±9.82%， 99.74%±12.05%， 58.91%±6.33%)， cell apoptosis rate increased(4.81%±0.26%， 4.93%±0.34%， 20.58%±2.11%）， cell migration(225.36±12.84， 224.89±19.56， 140.89±13.64) and invasion number(180.47±16.32， 177.54±6.92， 110.44±10.87) decreased， the expression levels of C-Caspase-9， E-cadherin and C-Caspase-3 proteins increased， the expression levels of N-cadherin， Notch1， Hes1 proteins decreased(P<0.05). Compared with the sh-ROR1-AS1 group， in the sh-ROR1-AS1+Jagged1 group， the survival rate increased(100.00%±11.58% vs 147.36%±12.05%）， cell apoptosis rate(19.54%±1.74% vs 8.36%±0.75%） decreased， cell migration(139.58±12.78 vs 175.26±16.94) and invasion numbers(107.45±9.35 vs 162.04±13.67) increased， the expression levels of C-Caspase-9， E-cadherin， C-Caspase-3 proteins decreased， and the expression levels of N-cadherin， Notch1， Hes1 proteins increased(P<0.05). ［Conclusion］ Down-regulation of ROR1-AS1 can inhibit cell proliferation， migration， invasion， EMT and activate cell apoptosis of breast cancer MCF-7 cells by inhibiting Notch1 signaling pathway. |
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