| 蒋可心,李宁,张旭.钙激活的氯离子通道 A4通过抑制 JAK激酶 2/信号转导及转录激活蛋白 3信号通路对食管癌细胞增殖、迁移及侵袭的影响[J].安徽医药,2021,25(3):474-478. |
| 钙激活的氯离子通道 A4通过抑制 JAK激酶 2/信号转导及转录激活蛋白 3信号通路对食管癌细胞增殖、迁移及侵袭的影响 |
| Effect of CLCA4 on proliferation, migration and invasion of esophageal cancer cells by inhibiting JAK2/STAT3 signaling pathway |
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| DOI：10.3969/j.issn.1009-6469.2021.03.010 |
| 中文关键词: 食管肿瘤 Janus激酶 2 STAT3转录因子 氯离子通道 A4(CLCA4) JAK2/STAT3信号通路 增殖 迁移 侵袭 |
| 英文关键词: Esophageal neoplasms Janus kinase 2 STAT3 transcription factor CLCA4 JAK2/STAT3 signaling path? way Proliferation Migra tion Invasion |
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| 中文摘要: |
| 目的探讨钙激活的氯离子通道 A4(CLCA4)对食管癌细胞增殖、迁移、侵袭及 JAK激酶 2/信号转导及转录激活蛋白 3(JAK2/STAT3)信号通路的影响。方法实时荧光定量逆转录聚合酶链反应( qRT-PCR)与蛋白质印迹法( Western blotting)分别检测正常人食管鳞状上皮细胞与人食管癌细胞中 CLCA4的表达；将合成的 CLCA4过表达载体及其对照分别转染至食管癌细胞 Eca109，分别记作 CLCA4过表达( pcDNA-CLCA4)组、 CLCA4阴性对照( pcDNA-NC)组，并将未转染的细胞作为阴性对照(NC)组。四甲基偶氮唑盐微量酶反应比色法( MTT法)检测细胞增殖能力；细胞迁移实验( Transwell)检测细胞迁移及侵袭能力。 JAK2/STAT3信号通路激活剂 p-JAK2多肽对细胞增殖、迁移及侵袭的影响。 Western blotting检测细胞周期蛋白 D1(Cy? clinD1)、依赖性激酶抑制因子( P21)基质金属蛋白酶 2(MMP-2)基质金属蛋白酶 9(MMP-9)、 JAK2、STAT3、磷酸化 JAK激酶 2(p-JAK2)、磷酸化信号转导及转录激、活蛋白 3(p-STAT3)的表达、水平。结果与 Het-1A相比，人食管癌细胞 KYSE170、 Eca109、TE10中 CLCA4 mRNA及蛋白表达水平降低( P<0.05)；与 pcDNA-NC组比较， pcDNA-CLCA4组细胞存活率显著降低［( 52.16±11.41)%比( 99.57±13.49)%，P<0.05］，迁移细胞数［( 56.47±10.03)%比( 112.49±13.52)%］与侵袭细胞数［( 63.43±9.87)%比( 123.47±16.58)%］减少( P<0.05)CyclinD1、MMP-2、MMP-9、p-JAK2、p-STAT3的表达水平降低( P<0.05)，P21的表达水平升高( P<0.05)；激活 JAK2/STAT3信号通路，可逆转 CLCA4过表达对 Eca109细胞增殖、迁移及侵袭的抑制作用。结论 CL?CA4过表达可抑制食管癌细胞增殖、迁移及侵袭，其作用机制可能与抑制 JAK2/STAT3信号通路活化有关。 |
| 英文摘要: |
| Objective To explore the effects of calcium-activated chloride channel A4 (CLCA4) on the proliferation, migration, inva? sion and non-receptor tyrosine protein kinase 2/signal transducer and activator of transcription 3 (JAK2/STAT3) signaling pathway ofesophageal cancer cells.Methods qRT-PCR and Western blotting were used to detect the expression of CLCA4 in normal humanesophageal squamous cells and human esophageal cancer cells; the synthetic CLCA4 overexpression vector and its control were trans?fected into esophageal cancer cells ( Eca109), denoted as CLCA4 overexpression (pcDNA-CLCA4) group, CLCA4 negative control (pcDNA-NC) group, and untransfected cells as negative control (NC) group. Thiazolan (MTT) detected cell proliferation ability; cell mi?gration test (Transwell) detected cell migration and invasion ability. The effect of JAK2/STAT3 signaling pathway activator p-JAK2 polypeptide on cell proliferation, migration and invasion. Western blotting detected Cyclin D1 (CyclinD1), dependent kinase inhibitor(P21), matrix metalloproteinase 2 (MMP-2), matrix metalloproteinase 9 (MMP-9), non-receptor tyrosine protein kinase 2 (JAK2), signaltransducer and activator of transcription 3 (STAT3), phosphorylated tyrosine kinase 2 (p-JAK2), phosphorylated signal transducer and activator of transcription 3 (p-STAT3). Results Compared with Het-1A, the expression levels of CLCA4 mRNA and protein in humanesophageal cancer cells KYSE170, Eca109 and TE10 were significantly lower (P<0.05). Compared with the pcDNA-NC group, the cell viability of the pcDNA-CLCA4 group was significantly decreased [(52.16±11.41)% vs. (99.57±13.49)%,P<0.05], and the number of mi? grated cells [(56.47±10.03)% vs. (112.49±13.52)%] and the number of invasive cells [(63.43±9.87)% vs. (123.47±16.58)%] were signifi? cantly decreased (P<0.05), the expression levels of CyclinD1, MMP-2, MMP-9, p-JAK2, p-STAT3 were significantly decreased (P< 0.05), and the expression level of P21 was significantly increased (P<0.05). Activation of the JAK2/STAT3 signaling pathway reversedthe inhibitory effect of CLCA4 overexpression on proliferation, migration and invasion of Eca109 cells. Conclusion Overexpression ofCLCA4 can inhibit the proliferation, migration and invasion of esophageal cancer cells, and its mechanism may be related to the inhibi?tion of JAK2/STAT3 signaling pathway activation. |
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