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| Study of the Effect of HOXC10 on the Stem Cell-Like Properties of Lung Cancer Cells Through Targeted Regulation of CST1 Expression |
| KANG Jianjun, GAO Le, XI Junfeng |
| The First Hospital of Yulin City, Shaanxi Suide 718000, China |
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Abstract Objective: To investigate the effect of HOXC10 on the stem cell-like characteristics of lung cancer cells through targeted regulation of CST1 expression. Methods: TLung cancer cell line A549 was cultured under serum-free conditions to form tumour spheres.WB detected the expression of HOXC10 and stemness marker proteins CD44,CD133,Nanog,SOX2 and OCT4 respectively.ShNC and ShHOXC10 A549 cells were constructed to detect the expression of stemness marker proteins CD44,CD133,Nanog,SOX2 and OCT4,tumour sphere-forming assay to detect cell self-renewal ability and CCK8 assay to detect cell proliferation ability.ShNC and ShHOXC10 lung cancer mouse subcutaneous tumour models were constructed,tumour volume and weight were measured,and the expression of HOXC10,CST1 and stemness marker proteins CD44 and CD133 were detected in tumour tissues by immunohistochemistry.CST1 mRNA and protein expression in A549 cells of ShNC and ShHOXC10 groups were detected by qPCR and WB.ChIP-PCR and ChIP-qPCR were used to detect HOXC10 and CST1 promoter sequence binding,and dual luciferase reporter genes were used to verify the regulation of CST1 promoter activity by HOXC10.The effect of CST1 overexpression on the expression of stemness marker proteins CD44,CD133,Nanog,SOX2,OCT4 and tumour spheroid formation and cell proliferation was examined by back-complementing pcDNA3.1-CST1 in ShHOXC10 cells. Results: HOXC10 was highly expressed in the tumour spheres of A549 cells,and knockdown of HOXC10 significantly inhibited the expression of stemness protein markers CD44,CD133,Nanog,SOX2,and OCT4,inhibiting the proliferation and tumour sphere-forming ability of A549 cells.Subcutaneous tumour assays showed that knockdown of HOXC10 inhibited the tumour-forming ability of A549 cells (P<0.001) and tumour weight (P<0.0001) and volume (P<0.01) were significantly reduced. Conclusion: HOXC10 regulates the stem cell-like properties of lung cancer cells through CST1.
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2022, Vol. 28 
