中国普外基础与临床杂志

中国普外基础与临床杂志

泛素特异性蛋白酶 39 对人结直肠癌细胞增殖的调控研究

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目的 观察结直肠癌(CRC)组织中泛素特异性蛋白酶 39(USP39)蛋白的表达情况,以及 USP39 基因敲低对结直肠癌细胞系 SW1116 和 HCT116 细胞生长和细胞周期的影响。 方法 ① 采用免疫组织化学染色方法检测 USP39 蛋白在结直肠癌组织的表达情况。② 选取结直肠癌 SW1116 和 HCT116 细胞系作为研究对象,将细胞分为 4 组(每组 5 个复孔):KD-1 组和 KD-2 组,采用慢病毒短发夹 RNA(shRNA)敲低肿瘤细胞中 USP39 基因的表达;shCon 组细胞感染阴性对照慢病毒,Con 组细胞未接受任何处理。采用四唑盐法(MTT)检测细胞的增殖能力,采用流式细胞仪检测细胞的周期分布。 结果 ① 免疫组织化学染色结果显示,USP39 蛋白在 CRC 组织中的高表达率高于癌旁组织(P=0.035)。② 不管是在 SW1116 细胞还是在 HCT116 细胞中,同时点下 KD-1 组和 KD-2 组细胞的增殖能力均明显低于 shCon 组和 Con 组(P<0.05)。③ 不管是在 SW1116 还是在 HCT116 细胞的 KD-1 组中,G0/G1 期的细胞百分比均较 Con 和 shcon 组降低(P<0.05),G2/M 期的细胞百分比和 sub-G1 期细胞数均增加(P<0.05)。 结论 USP39 高表达于 CRC 组织中。CRC 细胞系中敲低 USP39 基因的表达可抑制肿瘤细胞的增殖形成能力,促进肿瘤细胞早期发生凋亡。

Objective This study was aimed to determine the expression of ubiquitin-specific peptidase 39 (USP39) protein in the colorectal cancer (CRC) tissue, and the effect of silencing USP39 gene on the cell growth, colony formation, and cell cycle distribution of CRC cells. Methods ①The expressions of USP39 protein in CRC tissues and its paracancerous tissues were determined by immunohistochemical staining method. ②By lentiviral infection, Lv-shUSP39 (KD-1 and KD-2 group) and Lv-shCon (shCon group) were transferred into SW1116 and HCT116 cells, and cells of blank control group did not received any treatment (Con group). To determine the role of USP39 gene in cell growth, MTT assay was performed to draw growth curve, and cell cycle distribution of CRC cells in 4 groups were determined by flow cytometer. Results ① The expression of USP39 protein was higher in CRC tissues compared to adjacent tissues (P=0.035). ② For SW1116 and HCT116 cells, the cell proliferation ability of KD-1 and KD-2 groups were remarkably decreased than those in corresponding shCon and Con groups at the same time point (P<0.05). ③ Flow cytometry assay showed that, the percentage of G0/G1 phase cells were decreased obviously in G0/G1 (P<0.05), while increased significantly in percentage of G2/M phase and number of sub-G1 phase cells in KD-1 group compared with that in the Con gourp and shCon group of SW1116 and HCT116 cells (P<0.05). Conclusions The expression of USP39 protein was highly expressed in CRC tissues. Knockdowning of USP39 gene can inhibit cell proliferation and promoted cell apoptosis.

关键词: 结直肠癌; 泛素特异性蛋白酶 39; 短发夹 RNA; 细胞凋亡; 靶向治疗

Key words: colorectal cancer; ubiquitin-specific peptidase 39; shRNA; cell apoptosis; targeted therapy

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