Hela细胞诱导下TCR Vβ基因的选择性表达与重排研究

Study of the expression and rearrangement of TCR Vβ gene induced by Hela cells

目的研究Hela细胞诱导下T细胞受体(Tcell receptor,TCR)Vβ亚家族基因的优势表达规律,并探讨其是否与重排激活基因(RAGs)介导的TCR重排相关。方法将Hela细胞与健康人外周血单个核细胞(PBMC)混合培养,利用反转录聚合酶链式反应(reverse transcription-PCR,RT-PCR)检测共培养前后TCRVβ各亚家族基因、重排激活酶RAGs、非同源末端连接(non-homogousend joining,NHEJ)途径中的核心蛋白Ku70/Ku80及末端脱氧核苷酸转移酶(terminal deoxynucleotidyl transferase,TdT)等的表达特点。结果随着混合培养时间的推移,TCRVβ各亚家族基因由最初的全部表达逐渐转变为选择性的部分表达,其中,TCRVβ7在各时间段都有明显的表达;Ku70随着培养时间的推移表达量逐渐减少直至没有表达;而RAGs、ku80和TdT基因在各个时间段均未检测到其表达。结论在Hela细胞诱导下,TCRVβ各亚家族基因的表达有一定的选择性但无明显的特异性,该选择性表达可能与RAGs介导的二次重排无关。

Objective To study the expression pattern of TCR Vβ gene induced by Hela cells,and explore the correlation between the expression pattern and the rearrangement in T cell receptor（TCR） mediated by recombination activating genes（RAGs）.Methods The peripheral blood mononuclear cells（PBMC） and Hela cells were co-cultured.The expression of TCR Vβ,RAGs,Ku70/Ku80 in non-homologous end joining（NHEJ） pathway and the terminal deoxynucleotidyl transferase（TdT） genes were analyzed by reverse transcription polymerase chain reaction（RT-PCR）.Results All of TCR Vβ subfamily genes expressed equally before two cells co-cultured.When extend the culture time,the expression of TCR Vβ subfamily genes changed in different ways.TCR Vβ7 gene was highly expressed at all time,but the expression of Ku70 was gradually decreased until to no detection.Meanwhile,RAGs,ku80 and TdT genes were not detected.Conclusion The expression of TCR Vβ subfamily genes is only selective,no specific,when induced by Hela cells.The selective expression may be independent on RAGs-mediated secondary rearrangement.