Zhu Deqing, Li Xuan, Gong Hao, Li Jing, Lu Xike, Xia Honggang, Chen Xia, Ma Lan, Sun Zhongyi, Zhang Xun, Wang Dongbin
Department of Cardio-Thoracic Surgery, Tianjin Hospital, Tianjin 300211, China.
Institute of Clinical Orthopedics, Tianjin Medical University, Tianjin 300070, China.
Iran J Public Health. 2021 Apr;50(4):710-720. doi: 10.18502/ijph.v50i4.5995.
The effects of transthyretin (TTR) over-expression on the proliferation and cell cycle of non-small cell lung cancer (NSCLC) A549 cells and its possible mechanism were verified.
A total of 196 LC patients and 20 healthy controls were enrolled at Tianjin Hospital, Tianjin, China between Apr 2017 and Oct 2017. The serum TTR content was detected by ELISA. Through lentiviral transfection method, NSCLC cells were divided into non-transfected group (group A), negative control group (group B) transfected with empty vector and experimental group (group C) transfected with TTR over-expression. Cell proliferation was detected by CCK-8 method, TTR mRNA expression was detected by real-time quantitative polymerase chain reaction (RT-qPCR), and TTR protein expression was tested by Western blot (WB). Cell cycle was detected by flow cytometry, Wnt3a/β-catenin protein expression was detected by WB, and mRNA expression was detected by RT-qPCR.
The serum TTR content in early, middle and late LC group was remarkably lower than that in healthy group (<0.05). Compared with late stage, TTR content in early and middle stages of LC group was higher, and the difference was statistically marked ( < 0.05). The absorbance value of group C was lower than that of groups A and B, indicating that the cell proliferation activity dramatically decreased, with statistically marked difference (<0.05). LC A549 cells in group C were obviously blocked in G2M, with statistical significance (<0.05).
TTR over-expression can inhibit the proliferation of NSCLC A549 cells, and the expression is related to Wnt3a/β-catenin pathway. TTR in serum of patients was helpful for diagnosing LC and has certain clinical value.
验证转甲状腺素蛋白(TTR)过表达对非小细胞肺癌(NSCLC)A549细胞增殖和细胞周期的影响及其可能机制。
2017年4月至2017年10月期间,在中国天津医院招募了196例肺癌患者和20例健康对照。采用酶联免疫吸附测定法(ELISA)检测血清TTR含量。通过慢病毒转染法,将NSCLC细胞分为未转染组(A组)、转染空载体的阴性对照组(B组)和转染TTR过表达载体的实验组(C组)。采用CCK-8法检测细胞增殖,采用实时定量聚合酶链反应(RT-qPCR)检测TTR mRNA表达,采用蛋白质免疫印迹法(WB)检测TTR蛋白表达。采用流式细胞术检测细胞周期,采用WB检测Wnt3a/β-连环蛋白蛋白表达,采用RT-qPCR检测mRNA表达。
肺癌早、中、晚期组血清TTR含量均显著低于健康组(<0.05)。与晚期相比,肺癌组早、中期TTR含量较高,差异有统计学意义(<0.05)。C组吸光度值低于A组和B组,表明细胞增殖活性显著降低,差异有统计学意义(<0.05)。C组NSCLC A549细胞明显阻滞于G2M期,差异有统计学意义(<0.05)。
TTR过表达可抑制NSCLC A549细胞增殖,其表达与Wnt3a/β-连环蛋白通路有关。患者血清中的TTR有助于肺癌的诊断,具有一定的临床价值。