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目的：分析富含半胱氨酸的酸性分泌蛋白类似蛋白1（SPARCL1）对非小细胞肺癌（NSCLC）细胞增殖、凋亡、侵袭的影响，并探讨分裂原活化抑制剂（MEK）/细胞外调节蛋白激酶（ERK）通路在其中发挥的作用。方法：收集2019年9月~2021年6月期间本院接受手术治疗的84例NSCLC患者癌组织与相应癌旁组织，实时定量逆转录聚合酶链反应（qRT-PCR）法测定并比较各组织以及正常肺上皮细胞HBEpiC、NSCLC细胞A549、HCC827、H1299、H292中SPARCL1 信使RNA（mRNA）表达水平，选取A549、HCC827培养并分组，分为对照组、NC siRNA组、SPARCL1 siRNA组、U0126组（MEK/ERK特异性抑制剂）、SPARCL1 siRNA+U0126组，细胞计数法（CCK8）以及平板克隆法测定A549、HCC827细胞增殖，流式细胞仪测定A549、HCC827细胞凋亡，Transwell小室法测定A549、HCC827细胞侵袭能力，蛋白质印迹法（western blot）检测SPARCL1、p-MEK、MEK、p-ERK1/2、ERK1/2蛋白表达。结果：SPARCL1在NSCLC组织中mRNA表达水平低于癌旁组织（P＜0.05）；与HBEpiC细胞相比，NSCLC细胞A549、HCC827、H1299、H292细胞中SPARCL1 mRNA表达水平降低（P＜0.05）；与对照组相比，SPARCL1 siRNA组A549、HCC827细胞SPARCL1 mRNA表达水平与蛋白表达、凋亡率降低（P＜0.05），OD450、克隆形成数、侵袭细胞数、p-MEK/MEK、p-ERK1/2/ERK1/2蛋白表达升高（P＜0.05），U0126组A549、HCC827细胞SPARCL1 mRNA表达水平与蛋白表达、凋亡率升高（P＜0.05），OD450、克隆形成数、侵袭细胞数、p-MEK/MEK、p-ERK1/2/ERK1/2蛋白表达降低（P＜0.05）；与SPARCL1 siRNA组相比，SPARCL1 siRNA+U0126组A549、HCC827细胞SPARCL1 mRNA表达水平与蛋白表达、凋亡率升高（P＜0.05），OD450、克隆形成数、侵袭细胞数、p-MEK/MEK、p-ERK1/2/ERK1/2蛋白表达降低（P＜0.05）。结论：SPARCL1可能通过调控MEK/ERK通路影响NSCLC A549、HCC827细胞增殖、侵袭与凋亡。

英文摘要:

Objective: To analyze the influences of secreted protein acidic and rich in cysteines-like protein 1 (SPARCL1) on the proliferation, apoptosis and invasion of non-small cell lung cancer (NSCLC) cells, and to explore the role of Mitogen activation inhibitor (MEK) / Extracellular regulated protein kinase (ERK) pathway in it. Methods: From September 2019 to June 2021, the cancer tissues and corresponding adjacent tissues of 84 NSCLC patients who received surgical treatment in our hospital were collected, real time quantitative reverse transcription polymerase chain reaction (qRT-PCR) method was applied to measure and compare the expression level of SPARCL1 messenger RNA (mRNA) in tissues and normal lung epithelial cells HBEpiC and NSCLC cells A549, HCC827, H1299 and H292. A549 and HCC827 were selected for culture and grouping, they were divided into control group, NC siRNA group, SPARCL1 siRNA group, U0126 group (MEK/ERK specific inhibitor), and SPARCL1 siRNA+U0126 group. Cell counting kit 8 (CCK8) method and the plate cloning method were used to measure the proliferation of A549 and HCC827 cells, flow cytometry was applied to determine the apoptosis of A549 and HCC827 cells, Transwell chamber method was applied to determine the invasive ability of A549 and HCC827 cells, Western blot was applied to detect the protein expression of SPARCL1, p-MEK, MEK, p-ERK1/2, ERK1/2. Results: The SPARCL1 mRNA expression level in NSCLC tissues was lower than that in adjacent tissues (P<0.05). Compared with HBEpiC cells, the SPARCL1 mRNA expression level in NSCLC cells A549, HCC827, H1299 and H292 cells decreased (P<0.05). Compared with the control group, the SPARCL1 mRNA expression level, protein expression and apoptosis rate of A549 and HCC827 cells in SPARCL1 siRNA group decreased (P<0.05), the OD450, number of colonies formed, number of invasive cells, p-MEK/MEK, and p-ERK1/2/ERK1/2 protein expression increased (P<0.05), the SPARCL1 mRNA expression level, protein expression and apoptosis rate of A549 and HCC827 cells in the U0126 group increased (P<0.05), the OD450, number of colonies formed, number of invasive cells, p-MEK/MEK, and p-ERK1/2/ERK1/2 protein expression decreased (P<0.05). Compared with the SPARCL1 siRNA group, the SPARCL1 mRNA expression level, protein expression and apoptosis rate of A549 and HCC827 cells in SPARCL1 siRNA+U0126 group increased (P<0.05), the OD450, number of colonies formed, number of invasive cells, p-MEK/MEK, and p-ERK1/2/ERK1/2 protein expression decreased (P<0.05). Conclusion: SPARCL1 may affect the proliferation, invasion and apoptosis of A549 and HCC827 cells by regulating MEK/ERK pathway.

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