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目的 探究染色体不稳定(CIN)相关基因多肽N-乙酰半乳糖氨基转移酶7(GALNT7)对结肠癌细胞HCT116增殖和凋亡的影响。方法 采用慢病毒感染构建敲低GALNT7的HCT116细胞系，通过染色体滴定验证GALNT7与CIN的相关性，运用活细胞计数检测GALNT7对细胞增殖的影响，采用流式细胞术和Western blot检测GALNT7对细胞周期的影响，采用Caspase-3酶活性测定和Western blot检测GALNT7对细胞凋亡的影响。结果 敲低GALNT7后HCT116细胞染色体核型分布更为分散，CIN程度增高，细胞增殖速度减慢；敲低GALNT7后HCT116细胞发生细胞周期阻滞，并且细胞内P21蛋白上调，CDK6蛋白下调；敲低GALNT7后HCT116细胞Caspase-3酶活性上升，切割型PARP1蛋白和PUMA蛋白表达量升高，BCL-2蛋白表达量下降。结论 GALNT7基因可能通过抑制CIN产生促进结肠癌细胞增殖并抑制其凋亡。
Abstract：Objective To investigate the effect of chromosome instability(CIN) associated gene polypeptide N-acetylgalactosaminyltransferase 7(GALNT7) on proliferation and apoptosis of HCT116 colon cancer cells. Methods The HCT116 cell line with GALNT7 knockdown was constructed by lentiviral infection. The correlation between GALNT7 and CIN was verified by chromosome spread assay. The effect of GALNT7 on cell proliferation was detected by live cell counting, and the effect of GALNT7 on cell cycle distribution was detected by flow cytometry and Western blot. Caspase-3 activity and Western blot assays were used to detect the effect of GALNT7 on apoptosis. Results HCT116 cells showed a slower proliferation rate upon knocking down of GALNT7, and exhibited a more scattered karyotype distribution and a phenotype of increased degree of CIN. Inhibition of GALNT7 in HCT116 cells resulted in cell cycle arrest, upregulation of P21 and downregulation of CDK6 protein levels, as well as increased levels of Caspase-3 activity, cleaved PARP1 and PUMA protein expression, and decreased levels of BCL-2 protein expression. Conclusion The GALNT7 gene may promote proliferation and inhibit apoptosis of HCT116 colon cancer cells through the suppression of CIN generation.
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基本信息:
DOI：10.19405/j.cnki.issn1000-1492.2025.01.013
中图分类号:R735.35
引用信息:
[1]房晓,赵薇,喻文颖等.染色体不稳定相关基因GALNT7对结肠癌细胞HCT116增殖和凋亡的影响[J].安徽医科大学学报,2025,60(01):96-101.DOI:10.19405/j.cnki.issn1000-1492.2025.01.013.
基金信息:
国家自然科学基金(编号：81972251)~~