Skp2/p27轴在结直肠癌发生发展中的作用机制研究

doi:10.3969/j.issn.1005-6483.2021.11.017

摘要/Abstract

摘要： 目的探讨S期激酶相关蛋白2(Skp2)/p27轴参与结直肠癌(CRC)发生发展的分子机制，并初步研究其对CRC细胞LS513增殖、迁移侵袭及裸鼠成瘤能力的影响。
方法实时荧光定量聚合酶链式反应(qRT-PCR)检测Skp2和p27在CRC组织、癌旁组织以及人正常结肠上皮细胞CCD841CoN、不同CRC细胞株LS513、SW480、HT29和SW620中的表达。取生长密度达50%~60%的LS513癌细胞转染pSUPER-siRNA NC或pSUPER-siRNA Skp2，分别记为siRNA NC组和siRNA Skp2组，另取未转染LS513癌细胞(LS513组)。转染或培养24小时后，CCK-8法检测细胞增殖活性；Transwell小室法检测细胞迁移侵袭能力；裸鼠成瘤实验检测细胞在动物体内成瘤能力；蛋白免疫印迹(Western Blot)法检测细胞中Skp2、p27、p21、p57和CKS1蛋白表达水平。
结果与癌旁组织、CCD841CoN细胞相比，CRC组织、LS513、SW480、HT29和SW620癌细胞中Skp2 mRNA和蛋白表达水平显著升高，p27 mRNA和蛋白表达水平显著降低(P<0.05)。与LS513组、siRNA NC组相比，siRNA Skp2组细胞Skp2、CKS1蛋白表达水平、细胞增殖、迁移、侵袭及动物体内成瘤能力显著降低(P<0.05)，p27、p21、p57蛋白表达水平显著升高(P<0.05)。
结论 Skp2/p27轴参与CRC发生发展，干扰Skp2可增高CRC细胞中p27等抑癌蛋白表达，降低CKS1蛋白表达，抑制CRC细胞生长及迁移侵袭。

Abstract: Objective To investigate the molecular mechanism of Skp2/p27 axis involved in the occurrence and development of colorectal cancer(CRC) and to study its effect on the proliferation,migration and invasion of CRC cell line LS513 and tumorigenicity of nude mice.
Methods The expression of Skp2 and p27 in CRC tissues,paracancerous tissues,human normal colon epithelial cells CCD841CoN,different CRC cell lines LS513,SW480,HT29 and SW620 were detected by real-time fluorescence quantitative polymerase chain reaction(qRT-PCR).LS513 cells with a growth density of 50%-60% were transfected with siRNA NC or siRNA Skp2,respectively,and divided into siRNA NC group and siRNA Skp2 group respectively,in addition,LS513 cells without transfection(LS513 group) were taken.After 24 hours of transfection or culture,the cell proliferation was detected by CCK-8 method;Transwell chamber method was used to detect cell migration and invasion;the tumorigenicity of the cells in vivo was detected by nude mice tumorigenicity test;the protein expression levels of Skp2,p27,p21,p57 and CKS1 were detected by Western Blot.
Results Skp2 mRNA and protein expression levels in CRC tissues and LS513,SW480,HT29 and SW620 cancer cells were significantly higher than those in adjacent tissues and CCD841CoN cells,the expression levels of p27 mRNA and protein were significantly lower(P<0.05),Compared with those in LS513 group and siRNA NC group,the protein expression levels of Skp2 and CKS1,cell proliferation,migration,invasion and tumorigenicity in vivo were significantly lower in siRNA Skp2 group(P<0.05),and the protein expression levels of p27,p21 and p57 were significantly higher(P<0.05).
Conclusion Skp2/p27 axis is involved in the occurrence and development of CRC.Interference with Skp2 can increase the expression of tumor suppressor proteins such as p27 in CRC cells,decrease the expression of CKS1 protein,and inhibit the growth,migration and invasion of CRC cells.

Key words: S-phase kinase-associated protein 2/p27 axis, colorectal cancer, RNA interference, cell growth, migration and invasion

王海娟郑雅宾张静. Skp2/p27轴在结直肠癌发生发展中的作用机制研究[J]. 临床外科杂志, 2021, 29(11): 1054-1058.

WANG Haijuan, ZHENG Yabin, ZHANG Jing.. The role of Skp2/p27 axis in the occurrence and development of colorectal cancer[J]. JOURNAL OF CLINICAL SURGERY, 2021, 29(11): 1054-1058.

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