目的 探讨miR-567和钙周期素结合蛋白（CacyBP）对非小细胞肺癌（NSCLC）A549细胞增殖、侵袭和凋亡的影响机制。方法 选择miR-567、CacyBP高表达的NSCLC系A549作为研究对象,将实验分为4组,空白对照组（NC组）,转染pcDNA6.2 miR-567组（si-miR-567组）,转染pcDNA6.2 CacyBP siRNA组（si-CacyBP组）,转染pcDNA6.2 miR-567+CacyBP siRNA组（si-miR-567+CacyBP组）;qRT-PCR检测miR-567、CacyBP mRNA表达;CCK-8、MTT实验检测细胞增殖情况;Transwell实验检验细胞迁移、侵袭能力;流式细胞术检测NSCLC A549细胞凋亡率;Western bolt检测各组细胞凋亡蛋白变化情况。结果 相比于NC组,si-miR-567组、si-miR-567+CacyBP组miR-567表达显著降低;相比于NC组,si-CacyBP组、si-miR-567+CacyBP组CacyBP mRNA表达显著降低;MTT实验结果显示,三组细胞增殖抑制率均显著高于NC组,且si-miR-567+CacyBP组细胞增殖抑制率显著高于si-miR-567组、si-CacyBP组;CCK-8实验结果显示,三组细胞增殖计数均显著低于NC组,且si-miR-567+CacyBP组细胞增殖计数显著低于si-miR-567组、si-CacyBP组;Transwell实验结果显示NC组的细胞迁移、侵袭个数显著高于si-miR-567组、si-CacyBP组、si-miR-567+CacyBP组;且si-miR-567组、si-CacyBP组的细胞迁移、侵袭个数显著高于si-miR-567+CacyBP组;流式细胞术检测A549凋亡结果显示相比于NC组,si-miR-567组、si-CacyBP组、si-miR-567+CacyBP组的A549细胞凋亡率明显升高,且si-miR-567+CacyBP组的A549细胞凋亡率显著高于si-miR-567组、si-CacyBP组。结论 联合干预miR-567、CacyBP能够有效降低NSCLC A549细胞的增殖、迁移、侵袭能力,同时提高细胞的凋亡率。
Abstract
Objective To investigate the effects of miR-567 and calcyin-binding protein (CacyBP) on proliferation, invasion and apoptosis of non-small cell lung cancer (NSCLC) A549 cells. Methods NSCLC A549 with high expression of miR-567 and CacyBP was selected as the study object. The experiment was divided into 4 groups, blank control group (NC group), transfected with pcDNA6.2 miR-567(si-miR-567 group). Transfected pcDNA6.2 CacyBP siRNA (si-CacyBP group) and pcDNA6.2 miR-567+CacyBP siRNA group (si-miR-567+CacyBP group); mRNA expressions of miR-567 and CacyBP were detected by qRT-PCR. Cell proliferation was detected by CCK-8 and MTT assay. Transwell assay was used to test cell migration and invasion. The apoptosis rate of NSCLC A549 cells was determined by flow cytometry. The changes of apoptotic proteins in each group were detected by Western bolt. Results Compared with NC group, the expression of miR-567 in si-miR-567 groupand si-miR-567+CacyBP group was significantly decreased. Compared with NC group, the CacyBP mRNA expression in si-CacyBP group and si-miR-567+CacyBP group was significantly decreased. The results of MTT assay showed that the cell proliferation inhibition rate of three groups was significantly higher than that of NC group, and the cell proliferation inhibition rate of si-miR-567+CacyBP group was significantly higher than that of si-miR-567 and si-CacyBP groups. The results of CCK-8 experiment showed that the cell proliferation count of three groups was significantly lower than that of NC group, and the cell proliferation count of si-miR-567+CacyBP group was significantly lower than that of si-miR-567 and si-CacyBP groups. Transwell experiment showed that the number of cell migration and invasion in NC group was significantly higher than that in si-miR-567 group, si-CacyBP group and si-miR-567+CacyBP group. The number of cell migration and invasion in si-miR-567 group and si-CacyBP group was significantly higher than that in si-miR-567+CacyBP group. Flow cytometry showed that compared with NC group, the apoptosis rate of A549 cells in si-miR-567 group, si-CacyBP group and si-miR-567+CacyBP group was significantly increased. The apoptosis rate of A549 cells in si-miR-567+CacyBP group was significantly higher than that in si-miR-567 and si-CacyBP groups. Conclusion Combined intervention of miR-567 and CacyBP can effectively reduce the proliferation, migration and invasion ability of NSCLC A549 cells, and increase the apoptosis rate of cells.
关键词
钙周期素结合蛋白 /
miR-567 /
非小细胞肺癌 /
细胞增殖 /
细胞凋亡 /
分子机制
Key words
calcyclin binding protein /
miR-567 /
non-small cell lung cancer /
cell proliferation /
cell apoptosis /
molecular mechanism
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基金
河北省医学科学研究计划项目基金“miR-567对A549NSCLC细胞增殖和迁移的影响及其分子机制”（20221570）
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