Objective: To investigate the effect of centrosomal protein Cep63 on the apoptosis of papillary thyroid carcinoma (PTC) cell lines TPC-1 and underlying mechanism. Methods: With collected PTC tissues and adjacent tissues, Cep63 expression was detected by RT-qPCR and its relationship with clinicopathological factors was analyzed. The experiment included negative control group (NC), low expression group (Cep63(-)) and overexpression group (Cep63(+)), and wild-type TPC-1 cells were transfected with Cep63 lentivirus. The efficiency of Cep63 was detected by western blot (WB) and qRT-PCR. Cell proliferation ability was detected by plate cloning experiment and MTT assay. Cell apoptotic rate was detected by flow cytometry, and expression levels of apoptosis-related proteins were detected by immunohistochemistry and WB. The t-test was used to compare the differences in the means between the two groups, the one-way analysis of variance was used to compare multiple groups, and the chi-square test was used to analyze the association between gene expression levels and pathological factors. Results: Compared with NC group, cell proliferation ability was significantly decreased in Cep63(-) group (3.18±0.07 vs. 2.14±0.09, t=8.54, P<0.01) and significantly increased in Cep63(+) group (3.18±0.07 vs. 3.58±0.10, t=3.21, P<0.05). Apoptotic rates in NC, Cep63 (-) and Cep63 (+) groups were respectively 3.03%±0.24%, 8.66%±0.44% and 1.17%±0.44%, and the flow cytometry showed that the low expression of Cep63 significantly increased the apoptosis TPC-1 cells (F=157.7, P<0.001). Bcl-2 protein expression levels of NC, Cep63 (-) and Cep63 (+) groups were respectively 1.07±0.03, 0.49±0.01 and 1.99±0.09, and BAX protein expression levels of three groups were respectively 0.64±0.02, 1.06±0.01 and 0.21±0.03. WB showed that the expression level of Bcl-2 decreased (F=183.2, P<0.001), while the expression level of BAX was significantly up-regulated (F=283.7, P<0.001). Conclusion: Cep63 may regulate the apoptotic process of TPC-1 cells through Bcl-2/BAX pathway and Cep63 may be a potential oncogene of PTC.
目的： 研究中心体蛋白Cep63在甲状腺乳头状癌（papillary thyroid carcinoma，PTC）细胞系TPC-1凋亡过程中的作用及相关机制。 方法： 收集PTC病例的癌组织和癌旁组织，通过实时荧光定量PCR（qRT-PCR）检测组织Cep63的表达情况并分析其与临床病理因素的关系。本实验随机分为阴性对照组（NC）、低表达组［Cep63（-）］和过表达组［Cep63（+）］，将Cep63慢病毒转染野生型TPC-1细胞系，并通过蛋白免疫印迹法（WB）和qRT-PCR验证Cep63的干扰效果。通过平板克隆实验与MTT法检测细胞增殖能力变化；流式细胞术检测细胞的凋亡率，免疫组织化学（immunohistochemistry，IHC）和WB检测转染前后细胞凋亡相关蛋白表达差异。2组间均数比较采用t检验，单因素方差分析进行多组间均数差异的比较，病理因素关联分析采用卡方检验。 结果： 相比NC组，Cep63（-）组细胞增殖受到抑制（3.18±0.07比2.14±0.09，t=8.54，P<0.01），Cep63（+）组细胞的增殖能力明显提高（3.18±0.07比3.58±0.10，t=3.21，P<0.05）；NC组、Cep63（-）组和Cep63（+）组凋亡率分别为3.03%±0.24%、8.66%±0.44%和1.17%±0.44%，流式结果表明Cep63的低表达使TPC-1细胞的凋亡水平明显提高（F=157.7，P<0.001）；NC组、Cep63（-）组和Cep63（+）组Bcl-2表达量分别为1.07±0.03、0.49±0.01和1.99±0.09，BAX表达量分别为0.64±0.02、1.06±0.01和0.21±0.03，WB结果表明，Cep63的低表达使TPC-1细胞Bcl-2蛋白的表达量明显降低（F=183.2，P<0.001），同时BAX表达明显上调（F=283.7，P<0.001）。 结论： Cep63可能通过Bcl-2/BAX通路调控了PTC细胞系TPC-1的凋亡过程，Cep63可能是PTC的一个潜在癌基因。.