实用医学杂志 ›› 2020, Vol. 36 ›› Issue (24): 3345-3348.doi: 10.3969/j.issn.1006⁃5725.2020.24.008

• 基础研究 • 上一篇    下一篇

沉默TIPE 基因下调Wnt/β⁃catenin 信号对非小细胞肺癌细胞增殖、凋亡的影响

才德吉,措毛吉,万玛南杰

  

  1. 青海红十字医院 1 呼吸内科,2 产科,3 小儿外科(西宁 810000)

  • 出版日期:2020-12-25 发布日期:2021-01-19

The effect of silencing TIPE gene to down⁃regulating Wnt/β⁃catenin signal on proliferation and apoptosis of NSCLC cells

CAI Deji,CUO Maoji,WANMA Nanjie   

  1. Respiratory Medicine,Qinghai Red Cross Hospital Qin⁃ghai Xining 810000,China
  • Online:2020-12-25 Published:2021-01-19

摘要:

目的 研究沉默 TIPE(tumor necrosis factor⁃α⁃induced protein 8,TNFAIP8)基因下调 Wnt/β⁃ catenin信号对非小细胞肺癌细胞增殖、凋亡的影响。方法 通过构建shRNA⁃TIPE慢病毒载体转染人非小细 胞肺癌细胞系 A549,利用 Real time PCR、Western blot 检测 A549 TIPE、β⁃catenin 的表达水平,细胞克隆形 成实验、CCK8 检测细胞增殖,流式细胞仪检测细胞凋亡。结果 shRNA⁃TIPE 慢病毒转染非小细胞肺癌细 胞系A549后:(1)TIPE和β⁃catenin的mRNA、蛋白表达水平均低于阴性对照组和空白对照组;(2)TIPE基因沉默 后,A549细胞克隆形成能力显著下降(F = 17.48,P = 0.0003),生长曲线实验检测A549细胞增殖能力下降; 3)shRNA⁃TIPE慢病毒转染后,A549细胞凋亡率(18.4 ± 4.5)%明显高于阴性对照(12.7 ± 3.6)%和空白对照 组(11.5 ± 3.4)%,差异有统计学意义(F = 4.484 4,P = 0.035 1)。结论 TIPE可能通过激活Wnt/β⁃catenin 号通路促进非小细胞肺癌细胞系A549

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Abstract:

Objective To study the effect of silencing TIPE gene on Wnt/β⁃catenin signal on proliferation and apoptosis of NSCLC cells. Methods Human non ⁃small cell lung cancer cell line A549 was transfected with shRNA⁃TIPE by lentivirus vector. The expression levels of TIPE and β⁃catenin in A549 were detected by real time PCR and Western blot. Cell clone formation experiment,CCK8 and flow cytometry were used to detect cell prolifer⁃ ation and cell apoptosis. Results After transfection of shRNA⁃TIPE lentivirus was transfected into non⁃small cell lung cancer cell line A549,(1)The mRNA and protein expression levels of TIPE and β⁃catenin were lower than those of negative control group and blank control group;(2)The ability of clone formation of A549 cells decreased significantly after silencing of TIPE gene(F = 17.48,P = 0.000 3);(3)After transfection of shRNA⁃TIPE lentivirus the apoptotic rate(18.4 ± 4.5)% was significantly higher than that of negative control(12.7 ± 3.6)% and the blank control(11.5 ± 3.4)%,with statistical difference(F = 4.484 4,P = 0.035 1). Conclusion TIPE gene can promote proliferation and inhibit apoptosis of non⁃small cell lung cancer cell line A549 by activating Wnt/β⁃catenin 

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