| 马耀增,张茂锋,刘洪波,等.慢病毒介导的性别决定区 Y框蛋白 9表达下调对抑制 Wnt信号通路调控乳腺癌细胞侵袭能力的影响[J].安徽医药,2020,24(2):373-378. |
| 慢病毒介导的性别决定区 Y框蛋白 9表达下调对抑制 Wnt信号通路调控乳腺癌细胞侵袭能力的影响 |
| Inhibiting effect of down?regulation of lentivirus mediated sex dtermining region Y?box 9 expression on Wnt signaling pathway mediated invasion of breast cancer cells |
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| DOI：10.3969/j.issn.1009?6469.2020.02.042 |
| 中文关键词: 乳腺肿瘤 SOX9转录因子类 肿瘤浸润 Wnt信号通路 基质金属蛋白酶 2 钙黏着糖蛋白类 |
| 英文关键词: Breast neoplasms SOX9 transcription factor Neoplasm invasiveness Wnt signaling pathway Matrix metallo? proteinase 2 Cadherins |
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| 中文摘要: |
| 目的研究慢病毒介导的性别决定区 Y框蛋白 9(SOX9)表达下调对抑制 Wnt信号通路调控乳腺癌细胞侵袭能力的影响。方法以 Wnt信号通路抑制剂 XAV939处理乳腺癌细胞 MDA?MB?436，噻唑蓝( MTT)法测定细胞增殖活性，蛋白质免疫印迹(Western blot)检测细胞中 SOX9蛋白及 Wnt信号通路蛋白 β?连环蛋白( β?catenin)、下游靶基因 c?myc表达水平。 SOX9 siRNA慢病毒感染 MDA?MB?436细胞， Realtime PCR和蛋白质印迹法检测干扰效果。细胞划痕实验测定细胞迁移能力， Transwell小室检测细胞侵袭能力，蛋白质印迹法检测细胞中基质金属蛋白酶 ?2(MMP?2)和上皮间质转化( EMT)相关蛋白上皮性钙黏附素(E?cadherin)、神经性钙黏附素( N?cadherin)表达水平。结果 XAV939能够下调 MDA?MB?436细胞增殖活性，抑制乳腺癌细胞中 SOX9蛋白表达，降低 Wnt信号通路蛋白 β?catenin、c?myc表达水平。 SOX9 siRNA慢病毒感染后的 MDA?MB?436细胞中 SOX9 mRNA和蛋白水平降低。下调 SOX9的 MDA?MB?436细胞侵袭数目从( 85.56±4.35)降低至( 64.15±3.25)迁移率从(74.16±6.25)%降低至( 54.12±3.96)%，细胞中 MMP?2蛋白水平降低， E?cadherin蛋白水平升高， N?cadherin蛋白水，平降低。 XAV939也能够下调 MDA?MB?436细胞侵袭数目从( 85.56±4.35)降低至( 51.23±2.87)迁移率从( 74.16±6.25)%降低至( 45.28± 3.59)%，抑制细胞中 MMP?2蛋白表达，促进 E?cadherin蛋白表达，下调 N?cadherin蛋白表，达水平。 XAV939处理下调 SOX9的 MDA?MB?436细胞，细胞侵袭数目降低至( 34.18±2.59)迁移率降低至( 24.78±1.68)%，细胞中 MMP?2蛋白表达水平下降更多， E?cadherin蛋白水平升高更多， N?cadherin蛋白水平下多。结论抑制 Wnt信号通路下调乳腺癌细胞中 SOX9的表达水平，降更，敲低其表达能够协同 Wnt信号通路抑制剂降低乳腺癌细胞侵袭迁移能力。 |
| 英文摘要: |
| Objective To study the role of lentivirus mediated SOX9 down?regulation in the invasion of breast cancer cells by in?hibiting Wnt signaling pathway.Methods Wnt signaling pathway inhibitor XAV939 was used to treat breast cancer cell line MDA?MB?436.MTT assay was used to determine cell proliferation activity.Western blot was used to detect the expression level of SOX9protein and Wnt signaling pathway protein β?catenin and c?myc in cells.SOX9 siRNA lentivirus infection MDA?MB?436 cells，Real? time PCR and Western blot were used to detect the interference effect.Cell migration ability was measured by cell scratch test.Tran?swell was used to detect cell invasiveness.Western blot was used to detect the expression levels of MMP?2 and EMT related pro?teins E?cadherin and N?cadherin in cells.Results XAV939 can reduce the proliferation activity of MDA?MB?436 cells，inhibit SOX9 protein expression in breast cancer cells，and reduce the expression level of Wnt signaling pathway β?catenin and c?myc.Af? ter SOX9 siRNA lentivirus infection，the level of SOX9 mRNA and protein decreased in MDA?MB?436 cells，the number of inva? sive cells decreased from(85.56±4.35) to(64.15±3.25)， and the migration rate decreased from(74.16±6.25)% to(54.12±3.96)%，the level of MMP?2 protein in the cells decreased，the level of E?cadherin protein elevated，the level of N?cadherin protein decreased.XAV939 can also reduce the invasion and migration ability of MDA?MB?436 cells，the number of invasive cells de? creased from(85.56±4.35)to(51.23±2.87)and the migration rate decreased from(74.16±6.25)% to(45.28±3.59)%，and inhibit? ing the expression of MMP?2 protein in cellsromoted the expression of E?cadherin protein，and down?regulated the expression of N ，p?cadherin protein.XAV939 treatment of MDA?MB?436 cells decrease SOX9，the number of cell invasion was decreased to(34.18±2.59)，and the migration rate was decreased to(24.78±1.68)%.The expression level of MMP?2 protein in the cells decreased more， the level of E?cadherin protein increased more，the level of N?cadherin protein decreased more.Conclusion Inhibition of Wnt sig? naling pathway down?regulates SOX9 expression in breast cancer cells，knocking down its expression synergy with Wnt signaling pathway inhibitors reduces invasion and migration of breast cancer cells. |
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