富含亮氨酸重复序列和免疫球蛋白样结构域基因1过表达对膀胱癌细胞免疫相关基因的影响

Effect of leucine - rich repeats and immunoglobulin - like domains 1 gene overexpression on immu- nity - relative genes in bladder cancer cells

目的利用基因表达谱芯片技术筛选过表达富含亮氨酸重复序列和免疫球蛋白样结构域基因1（LRIG1）的膀胱癌细胞与对照组膀胱癌细胞间差异表达的免疫相关基因，探讨LRIG1对膀胱癌细胞免疫功能影响的机制。方法分别提取转染LRIG1基因的膀胱癌细胞BIU87-LRIG1和未转染LRIG1基因的膀胱癌细胞BIU87的总核糖核酸（RNA），反转录成cDNA并标记后，同22K人类基因表达谱芯片杂交后筛选出与免疫相关的差异表达基因，并进一步使用反转录-聚合酶链反应（RT—PCR）验证。结果在21522条基因中，两组细胞间差异表达的免疫相关基因有9条，其中BIU87-LRIG1细胞中上调基因4条，下调基因5条。RT-PCR提示BIU87-LRIG1组的程序化死亡基因5（PDCD5）表达水平（1．22±0．04）较BIU87组（0．43±0．06）明显增高。结论使用基因芯片技术筛选出了LRIG1过表达可影响膀胱癌免疫功能的9种相关基因。

Objective To investigate the effects of leucine - rich repeats and immunoglobulin - like domains 1 （LRIG1） gene overexpression on immunity- relative genes in bladder cancer BIU87 cells by cDNA microarray technique and to explore the mechanism of LRIG1 affecting immune functions in blad- der cancer ceils. Methods The total RNA was extracted and purified from BIU87 cells transfected with LRIG! （BIU87 - LRIG1 cells） and control cells, respectively. The cDNA was obtained and labeled with fluorescence, and then hybridized with gene chip containing human 22K gene expression profile. Subse- quently, the signal images were scanned and analyzed. The reverse transcription- polymerase chain reac- tion （ RT - PCR） was used to confirm the array results. Results Of the 21 522 target genes, 9 genes were screened out for differences in immunity - relative gene expression level. Four of the 9 genes were up - regulated and 5 were down - regulated in the BIU87 - LRIG1 cells. The results of RT - PCR indicated that the expression level of PDCD5 in the BIU87 - LRIG1 cells （ 1.22 ± 0. 04） was significantly higher than in control cells （0. 43 ± 0. 06）. Conclusion Nine immunity - relative genes in bladder cancer ceils are found by using gene chip.