文章摘要
过表达NOLC1诱导人非小细胞肺癌细胞凋亡
Overexpression of NOLC1 Induced Apoptosis in Human Non-Small Cell Lung Cancer Cells
  
DOI:doi:10.3969/j.issn.1005-7021.2024.02.009
中文关键词: NOLC1(Nucleolar and coiledbody phosphoprotein 1)  肺癌细胞  腺病毒载体  过表达  细胞凋亡
英文关键词: NOLC1  lung cancer cells  adenovirus  overexpression  apoptosis
基金项目:辽宁省教育厅一般项目(LJC201910);辽宁省教育厅服务地方项目(LJKFZ20220179)
作者单位
杜亚兰 辽宁大学 生命科学学院辽宁 沈阳 110000 
张茂盛 秦皇岛市海港医院河北 秦皇岛 066000 
王旌羽 辽宁大学 生命科学学院辽宁 沈阳 110000 
张璐燕 辽宁大学 生命科学学院辽宁 沈阳 110000 
周常博 辽宁大学 生命科学学院辽宁 沈阳 110000 
佘晓双 宁阳县第一人民医院 山东 泰安 271000 
郑方亮 辽宁大学 生命科学学院辽宁 沈阳 110000 
朱春玉 辽宁大学 生命科学学院辽宁 沈阳 110000 
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中文摘要:
      人核仁磷酸化蛋白1 (Nucleolar and coiledbody phosphoprotein 1,NOLC1)在癌症的发生发展过程中起着至关重要的调控作用,为探讨NOLC1对肺癌细胞的作用,本研究通过Gateway系统构建重组NOLC1腺病毒载体,成功包装NOLC1腺病毒后,分别感染正常人类胚胎肺细胞(HEL)和非小细胞肺癌细胞(A549细胞),过表达NOLC1。通过MTT实验、AnnexinV-APC/PI双染法和线粒体膜电位实验,证明与HEL细胞相比,NOLC1的过表达对A549细胞的活性降低、凋亡增加、线粒体膜电位下降影响较为显著;通过Real-time PCR检测Caspase家族、TNF与受体家族和BCL2家族基因的表达,发现过表达NOLC1明显上调了A549细胞中促凋亡基因的表达,下调了抗凋亡基因的表达,其中两种重要的促凋亡蛋白CASP8和BAX均显著上调,但是在HEL细胞中这种影响不明显。研究结果表明过表达NOLC1蛋白通过对线粒体通路和死亡受体通路的共同作用,对非小细胞癌具有显著的抗肿瘤活性。
英文摘要:
      Human NOLC1 (nucleolar and coiledbody phosphoprotein 1) play an imperative regulation role in the onset and progression of cancer. A recombinant NOLC1 adenovirus vector using the Gateway system was established in order to study the impact of NOLC1 on lung cancer cells. After successfully packaging the NOLC1 adenovirus, and respectively infected normal human embryonic lung cells (HEL) and non-small cell lung cancer cells (A549 cells), to overexpress the NOLC1. The overexpression of NOLC1 had more significant impact on reduction activity, apoptosis increment, and decrement of mitochondrial membrane potential in A549 cells.. Adoption of MTT assay, AnnexinV-APC/PI double-staining assay, and mitochondrial membrane potential assay; it was proved as compared with HEL cells the expression of Caspase family, TNF and receptor family, and BCL2 family genes were detected by Real-time PCR. Real-time PCR was used to measure the expression of genes from the Caspase family, TNF and receptor family, and BCL2 family. It was discovered that, in A549 cells, overexpression of NOLC1 significantly increased the expression of pro-apoptotic genes and decreased the expression of anti-apoptotic genes, among them two key pro-apoptotic proteins, CASP8 and BAX, being significantly up-regulated. However, this effect was not evident in HEL cells. The study results suggested that the overexpression of NOLC1 protein has significant anti-tumor activity against non-small cell carcinoma through a combined effect on the mitochondrial pathway and death receptor pathway.
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