活化诱导胞嘧啶核苷酸脱氨酶对膀胱癌T24细胞表型的影响

The effects of activation-induced cytosine deaminase on the phenotype of bladder cancer T24cells

目的探讨AID(活化诱导胞嘧啶核苷酸脱氨酶)在人体膀胱癌、癌旁组织中的表达情况以及AID对膀胱癌细胞株T24增殖、侵袭、迁移以及凋亡的影响。方法运用Western bolt以及免疫组织化学染色法检测16例临床膀胱癌组织以及癌旁组织中AID的相对表达量;将携带有抑制AID表达的shRNA序列(shAICDA-T24组)或空载序列(NC-T24组)以慢病毒为载体对T24细胞进行感染,利用Western bolt对T24、NC-T24以及shAICDA-T24组进行AID表达量的检测;因为转染后的多克隆细胞株AID的表达抑制效果有限,随后我们对转染的T24细胞进行单克隆细胞筛选,通过Western bolt结果选取抑制效果最明显的组别用于后续试验;然后运用细胞增值试验(CCK8)、流式细胞术(凋亡)、细胞划痕实验以及细胞小室实验(Transwell)检测抑制AID的表达对T24细胞增殖、凋亡、侵袭以及迁移的影响。结果人体膀胱癌组织中存在AID的表达,并且膀胱癌组织中AID的表达显著高于癌旁组织(P<0.05);通过RNA干扰技术(RNAi)抑制AID的表达可显著降低膀胱癌细胞株T24的增殖、侵袭和迁移能力,并且增加了T24细胞的凋亡率(P<0.05)。结论抑制AID的表达可显著降低T24细胞增殖、侵袭以及迁移能力,并且增加T24细胞的凋亡,其表达可能与膀胱癌的进展具有相关性。

Objective To investigate the expression of activation-induced cytosine deaminase （AID）in human bladder cancer and para-cancerous tissues and its role in bladder cancer cell line T24. Methods The relative expressions of AID in 16 cases of bladder cancer tissues were detected with Western bolt and immunehistochemical staining. The T24 cells were transfected with shRNA or no-load lentiviral vector and divided into shAICDA-T24 group and NC-24 group. The expressions of AID in T24 group,shAICDA-T24 group and NC-24 group were detected with Western bolt. As the inhibitory effects were not effective on polyclone cells, the monoclone cells were screened out. The effects of AID on the proliferation, apoptosis,invasion and migration of T24 cells were assessed with counting kit 8 （CCK8）, flow cytometry apoptosis assay, Transwell assay and wound healing assay. Results AID expression was significantly higher in bladder cancer tissues than in paracancerous tissues （P〈0.05）. The inhibition of AID expression by RANi could significantly reduce the proliferation,migration and invasion abilities of T24 cells （P〈0.05）, and increase the apoptosis of T24 cells. Conclusion The inhibition of AID expression can reduce the proliferation,migration and invasion of T24 cells but increase the apoptosis. AID expression may be related to the occurrence and progression of bladder cancer.