三结构域包含蛋白29对鼻咽癌细胞生长和迁移的影响

Effects of TRIM 29 on growth and migration of nasopharyngeal carcinoma cells

目的研究三结构域包含蛋白(TRIM 29)、死骨片蛋白-1(SQSTM 1)、根蛋白(RDX)基因在不同转移潜能鼻咽癌细胞系中的表达水平,了解其与鼻咽癌细胞侵袭转移的关系,揭示TRIM 29在鼻咽癌细胞侵袭转移中的作用。方法 1采用q RT-PCR(quantitative reverse transcriptase polymerase chain reaction)和蛋白印迹法(Western blot)检测高转移潜能鼻咽癌细胞系5-8F和低转移潜能鼻咽癌细胞系6-10B中TRIM 29、SQSTM 1、RDX基因的m RNA和蛋白表达水平;2脂质体转染法将TRIM 29特异性si RNA载体和空白载体转染5-8F细胞,建立稳定转染细胞系;3采用Western blot检测细胞中TRIM 29蛋白表达水平的变化,MTT法检测细胞增殖能力的变化,流式细胞仪检测细胞周期分布的变化,划痕实验观察细胞迁移能力的变化。结果 15-8F细胞中TRIM 29、SQSTM 1的表达水平明显高于6-10B细胞,而RDX的表达水平低于6-10B细胞;2建立了TRIM 29低表达的5-8F细胞系及其空白载体稳定转染细胞系;3与空白载体转染的5-8F细胞和未转染5-8F细胞比较,特异性si RNA转染的TRIM 29低表达5-8F细胞的增殖能力降低,G0/G1细胞增加,而S期细胞减少,细胞迁移能力降低。结论 1si RNA干扰TRIM 29表达抑制鼻咽癌5-8F细胞的生长和迁移;2TRIM 29、SQSTM 1与鼻咽癌转移密切相关,有望成为预警鼻咽癌转移的分子标志物,但RDX与鼻咽癌转移关系不确定。

【Objective】 To explore the expression of TRIM 29, SQSTM 1, RDX genes in nasopharyngeal carcinoma（NPC） cell lines with different potential metastatic ability so as to know the relationship between TRIM 29, SQSTM 1, RDX and the metastasis of NPC cells and to study the role of TRIM 29 in the invasion and metastasis of NPC cells. 【Methods】 1Quantitative reverse transcriptase polymerase chain reaction（q RTPCR） and Western blot were used to respectively detect the m RNA levels and the protein expression levels of TRIM 29, SQSTM 1, RDX in high metastatic NPC cell line（5-8F） and lower metastatic NPC cell line（6-10B）; 2Liposome transfection method was used to establish the stably transfected 5-8F cells with TRIM 29 specific small interfering RNA（Si RNA）, and its corresponding control cells with empty vector transfection; 3Western blot, MTT assay, flow cytometry and wound healing assay were performed to compare the differences in protein expression level, cell proliferation, cell cycle distribution and cell migration in the stably transfected NPC cells. 【Results】 1The m RNA levels and the protein expression levels of TRIM 29, SQSTM 1 in 5-8F cells were significantly higher than those in 6-10 B cells, whereas RDX expression level was lower than that in 6-10 B cells; 2The stable transfected 5-8F cell line with low TRIM 29 expression and its corresponding control cell line were established; 3Compared with empty vector transfected and untransfected 5-8F cells, the5-8F cells with TRIM 29 specific si RNA showed the decreased proliferation, increased percent of G0/ G1 phase cells, reduced percent of S phase cells, and decreased migration ability. 【Conclusion】 1TRIM 29 Si RNA inhibits the growth and migration ability of 5-8F cells; 2TRIM 29 and SQSTM 1 are closely related to the metastasis of NPC, may be the important molecular markers predicting metastasis of NPC, but the relationship between RDX and NPC is uncertain.