USP22 siRNA通过TIMP-2抑制人结肠癌细胞系SW480侵袭
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摘要
目的探讨泛素特异性蛋白酶22(USP22)调控基质金属蛋白酶组织抑制物-2(TIMP-2)对人结肠癌细胞侵袭的影响。方法 Western blot检测USP22在不同结肠癌细胞系的表达,选定高表达USP22的SW480细胞用于后续实验;将SW480细胞分为对照组(NC siRNA)和转染组;实时荧光定量PCR(RT-qPCR)检测对照组及转染组Hif-1α、Hif-1β、cdc42、RhoA、Smad7、TIMP-1和TIMP-2 mRNA表达;RT-qPCR和Western blot检测对照组及2种转染组中USP22及TIMP-2蛋白的表达;Transwell小室法检测细胞的侵袭能力。结果 USP22在SW480细胞中表达量较高(P<0.05);与对照组相比敲低USP22基因后,TIMP-2表达及SW480细胞侵袭能力均下降(P<0.05);敲低TIMP-2基因后,USP22表达无明显变化,SW480细胞侵袭能力下降(P<0.05)。结论 USP22可能通过调控TIMP-2的表达抑制人结肠癌细胞的侵袭与转移。
Objective To study the role of USP22 and its target-controlled TIMP-2 on invasion of human colonic cancer cells. Methods The expression of USP22 in colonic cancer cells was detected by Western blot. SW480 cells were used for further experiments because of its high expression. The SW480 cells were categoried into transfected group and control group(NC siRNA). The expression of Hif-1α,Hif-1β,Cdc42,RhoA,Smad7,TIMP-1,TIMP-2 mRNA was detected by quantitative real-time PCR(RT-qPCR) in control group and trasfected group. The expression of USP22 and TIMP-2 was determined by Western blot and RT-qPCR in control group and two transfected groups. Invasive activities of SW480 was detected by Transwell. Results USP22 expression was highly expressed in SW480(P<0.05). After silencing the USP22 gene, TIMP-2 expression and the invasive activity of SW480 were suppressed as compared with control group(P<0.05). Knockdown of TIMP-2 gene did not infuence the expression of USP22, but the invasive of SW480 was suppressed(P<0.05). ConclusionsUSP22 may inhibit the invasion and metastasis of colonic cancer by regulating the expression level of TIMP-2.
Objective To study the role of USP22 and its target-controlled TIMP-2 on invasion of human colonic cancer cells. Methods The expression of USP22 in colonic cancer cells was detected by Western blot. SW480 cells were used for further experiments because of its high expression. The SW480 cells were categoried into transfected group and control group(NC siRNA). The expression of Hif-1α,Hif-1β,Cdc42,RhoA,Smad7,TIMP-1,TIMP-2 mRNA was detected by quantitative real-time PCR(RT-qPCR) in control group and trasfected group. The expression of USP22 and TIMP-2 was determined by Western blot and RT-qPCR in control group and two transfected groups. Invasive activities of SW480 was detected by Transwell. Results USP22 expression was highly expressed in SW480(P<0.05). After silencing the USP22 gene, TIMP-2 expression and the invasive activity of SW480 were suppressed as compared with control group(P<0.05). Knockdown of TIMP-2 gene did not infuence the expression of USP22, but the invasive of SW480 was suppressed(P<0.05). ConclusionsUSP22 may inhibit the invasion and metastasis of colonic cancer by regulating the expression level of TIMP-2.
引文
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[4] Tarcic O, Granit RZ, Pateras IS, et al. RNF20 and histone H2B ubiquitylation exert opposing effects in Basal-Like versus luminal breast cancer [J]. Cell Death Differ, 2017, 24: 694-704.
[5] Wang Z, Zhu L, Guo T, et al. Decreased H2B monoubiquitination and overexpression of ubiquitin-specific protease enzyme 22 in malignant colon carcinoma [J]. Hum Pathol, 2015, 46: 1006-1014.
[6] He Y, Jin YJ, Zhang YH, et al. Ubiquitin-specific peptidase 22 overexpression may promote cancer progression and poor prognosis in human gastric carcinoma [J]. Transl Res, 2015, 165: 407-416.
[7] Ji M, Shi H, Xie Y, et al. Ubiquitin specific protease 22 promotes cell proliferation and tumor growth of epithelial ovarian cancer through synergy with transforming growth factor beta1 [J]. Oncol Rep, 2015, 33: 133-140.
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[11] Ao N, Wang L, Liu Y. Prognostic and clinicopathological significance of ubiquitin-specific protease 22 overexpres-sion in cancers: evidence from a meta-analysis [J]. Onco Targets Ther, 2017, 10:5533-5540.
[12] Ma Y, Fu HL, Wang Z, et al. USP22 maintains gastric cancer stem cell stemness and promotes gastric cancer progression by stabilizing BMI1 protein [J]. Oncotarget, 2017, 8: 33329-33342.
[13] Liu YL, Zheng J, Tang LJ, et al. The deubiquitinating enzyme activity of USP22 is necessary for regμLating HeLa cell growth [J]. Gene, 2015, 572: 49-56.
[14] Pen?e S, ?zbek E, Ozan Tiryakio variant genotype frequency and increased TIMP-2 and MMP-9 expression are positively correlated with cancer invasion in urinary bladder cancer[J]. Cell Mol Biol (Noisy-le-grand), 2017, 63: 46-52.