CD133^+皮肤鳞状细胞癌细胞的增殖与自我更新和转移能力

Proliferation,Self-renewal and Metastasis Ability of CD133^+ Cells in Cutaneous Squamous Cell Carcinoma

【目的】研究CD133阳性亚群在皮肤鳞状细胞癌(cSCC)的生物学特性,以探讨cSCC的肿瘤干细胞(CSC)自我更新和促进cSCC恶性行为的机制。【方法】采用流式分选皮肤鳞状细胞癌Colo-16细胞系中CD133^+(阳性群体)和CD133^-(阴性群体)细胞,应用四甲基偶氨唑盐检测两群细胞的增殖活性;体外成球实验检测二者自我更新能力的差异;Transwell法检测细胞浸润能力;应用Western Blot和荧光定量PCR方法检测CD44、SOX2等肿瘤干细胞特性(干性)相关基因在两群细胞中的表达情况;免疫组化了解日光性角化病、鲍温病和cSCC组织中CD133的表达情况。【结果】CD133^+Colo-16细胞表现出了显著更强的体外增殖、自我更新和浸润能力(P<0.05);CD133^+细胞中干性相关基因CD44、SOX2和OCT4的蛋白表达及对应的mRNA转录水平均高于CD133^-细胞(P<0.05);另外,CD133在日光性角化病、鲍温病和cSCC组织中表达呈逐步上调趋势。【结论】CSC的分子标记CD133的表达与肿瘤细胞的增殖、自我更新和浸润等恶性特征相关,CD133阳性cSCC细胞具有CSC干性。cSCC的发生发展可能与表达CD133阳性的CSC有关。

Abstract:【Objective】To observe the biological characteristics of CD133positive cells derived from cutaneous squamous cell carcinoma(cSCC),and to investigate the mechanism of self-renewal of cancer stem cells(CSC)and the malignant behavior in cSCC.【Methods】The flow cytometry was applied to purify CD133+cells and CD133-from cutaneous squamous cell carcinoma cell line Colo-16.The effects of CD133on the cell proliferative,self renew and migratory activity of two group cells were detected by MTT assay,Sphere formation assay and Transwell assay.Western blot and quantitative fluorescent PCR were performed to investigate the expression of cancer stem cell associated gene CD44,OCT4and SOX2in two groups of cells.Examined the expression level of CD133in actinic keratosis,squamous cell carcinoma in situ and cSCCby immunohistochemistry.【Result】The CD133+Colo-16cells exhibited significantly stronger proliferation,self-renewal and metastasis ability(P<0.05)and present higher level protein and mRNA expression on CD44,SOX2and OCT4(P<0.05).In addition,the expression level of CD133was dynamic rise during the continuous evolution of actinic keratosis,squamous cell carcinoma in situ and cSCC.【Conclusions】The expression level of CD133may be related to the tumorigenesis of cSCCand the malignant behavior including proliferative,self renew and migratory activity,and CD133+cSCC cells show the properties of cancer stem cells.The occurrence and development of cSCC may be related to the CSC,which express CD133.