| 王红,王恳,焦庆昉,等.YAP干扰腺病毒载体对宫颈癌增殖、迁移及侵袭的影响[J].安徽医药,2021,25(6):1136-1139. |
| YAP干扰腺病毒载体对宫颈癌增殖、迁移及侵袭的影响 |
| Effect of adenovirus-mediated silencing of YAP on the proliferation, migration, and invasion of cervical carcinoma |
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| DOI：10.3969/j.issn.1009-6469.2021.06.018 |
| 中文关键词: 子宫肿瘤 YAP Hippo信号通路 CaSki细胞 恶性增殖 迁移 侵袭 |
| 英文关键词: Uterine neoplasms YAP Hippo signaling pathway CaSki cells Malignant proliferation Migration Invasion |
| 基金项目: |
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| 摘要点击次数: 1294 |
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| 中文摘要: |
| 目的探索 YAP蛋白对宫颈癌细胞的恶性增殖、迁移及侵袭性的作用。方法分别以腺病毒靶向 YAP干扰载体(pAd-si-YAP)、重组空载体腺病毒( pAd-mock)或磷酸缓冲盐溶液( PBS)感染 CaSki细胞。采用定量聚合酶链反应( qPCR)、蛋白质印迹法( Western blotting)、四甲基偶氮唑盐微量酶反应比色法( MTT法)、流式细胞法、划痕试验以及细胞侵袭试验分别分析感染后 CaSki细胞的 YAP mRNA表达、 YAP蛋白表达、细胞增殖活力、细胞周期及凋亡、细胞迁移能力以及细胞侵袭能力。结果 qPCR结果显示 Ad-si-YAP感染 CaSki细胞的 YAP mRNA表达明显低于 pAd-mock感染的 CaSki细胞及 PBS处理的 CaSki细胞［(22.01±0.24)比( 80.12±0.31)、(84.18±0.22)P<0.05］表明 Ad-si-YAP可以抑制 YAP基因转录。蛋白质印迹法结果显示 Ad-si-YAP感染CaSki细胞的YAP蛋白表达低于pA，d-mock感，染的 CaSki细胞及 PBS处理的 CaSki细胞［( 0.6±0.018)比( 1.5±0.031)、(1.8±0.27)P<0.05］。 MTT结果显示 24 h后 pAd-mock感染的 CaSki细胞及 PBS处理的 CaSki细胞增殖明显快于 Ad-siYAP感染 CaSki细P<0.05)。流式细胞结果显示通过沉默 YAP后， CaSki细胞周期停滞于 G0/G1期明显增加( P<0.05)同时 CaSki细胞增多( P<0.01)。划痕试验发现沉默 YAP后 CaSki细胞的迁移能力下降［(40.01±0.16)比( 81.02±0.22)、(86.04±01)， P<0.05］说明 YAP可以促进 CaSki细胞迁移。细胞侵袭试验发现沉默 YAP后 CaSki细胞的侵袭能力下降［(120±4)比( 640±3)、胞(.3，(680±4)，P<0.05］。结论 YAP蛋白的表达可导致宫颈癌细胞的恶性增殖、迁移及侵袭，是应用于宫颈癌综合治疗的一个潜在靶点。 |
| 英文摘要: |
| Objective To explore the effect of adenovirus-mediated YAP-silencing system (pAd-si-YAP) on the malignant proliferation, migration, and invasion of cervical carcinoma.Methods pAd-si-YAP, mock-recombinant adenovirus (pAd-mock), and phosphatebuffer saline (PBS) were transfected into CaSki cell lines. The YAP mRNA/protein expression level, the cell proliferative potential, thecell cycle and apoptosis, and the migration/invasive potential of the transfected CaSki cell lines were determined by using quantitativepolymerase chain reaction (qPCR), Western blotting, MTT assay, flow cytometry, wound healing assay, and Transwell assay, respectively.Results In this experiment, qPCR indicated that CaSki cells transfected with Ad-si-YAP showed a significantly lower YAP mRNA expression level compared to those transfected with pAd-mock or treated with PBS[(22.01±0.24) vs. (80.12±0.31), (84.18±0.22), P< 0.05]. Western blotting indicated that CaSki cells transfected with Ad-si-YAP showed a significantly lower YAP protein expression level compared to those transfected with pAd-mock or treated with PBS[(0.6±0.018) vs. (1.5±0.031), (1.8±0.27), P<0.05]. MTT assay indicated that CaSki cells transfected with pAd-mock or treated with PBS proliferated more rapidly compared to those transfected with Ad-si-YAP (P<0.05). Flow cytometry indicated that Yap silencing caused more CaSki cells to arrest at the G0/G1 phase (P<0.05) and resulted in a higher cell count (P<0.01). Wound healing assay revealed a decrease in the migration potential of the CaSki cells after YAP silencing[(40.01±0.16) vs. (81.02±0.22), (86.04±0.31), P<0.05]. Transwell assay revealed a decrease in the invasive potential of the CaSki cells after YAP silencing[(120±4) vs. (640±3), (680±4), P<0.05].Conclusions The expression of YAP protein can induce malignant proliferation, migration, and invasion of cervical carcinoma cells, and it is a potential target for the comprehensive treatment of cervical carcinoma. |
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