摘要
目的:研究ATP结合盒亚家族A成员8(ABCA8)对结直肠癌(CRC)细胞增殖、迁移与侵袭的影响,并阐明其作用机制。方法:采用生物信息学分析ABCA8在CRC组织中的表达情况;采用实时荧光定量PCR(RT-qPCR)法检测临床CRC癌组织和CRC细胞系中ABCA8 mRNA表达水平。采用ABCA8基因过表达慢病毒感染SW480细胞,再将细胞分为对照(Blank)组、空载(Vector)组、ABCA8过表达(Oe-ABCA8)组、Vector+HLY78组和Oe-ABCA8+HLY78组,其中Wnt/β-catenin信号通路激动剂HLY78干预浓度为10μmol/L。CCK-8法检测各组细胞增殖活性;Transwell实验检测各组细胞迁移与侵袭细胞数量;Western blot法检测各组细胞中ABCA8、β-catenin、c-Myc和金属基质蛋白酶9(MMP-9)蛋白表达水平。结果:ABCA8在CRC癌组织和CRC细胞系SW480、SW620、 HCT116和HT-29中均呈低表达水平。与Blank组比较,Oe-ABCA8组SW480细胞增殖活性显著降低(P<0.05),迁移与侵袭细胞数量显著减少(P<0.05),细胞中ABCA8表达显著升高(P<0.05),β-catenin、c-Myc和MMP-9蛋白表达水平均显著降低(P<0.05),而Vector组各指标无显著性差异(P>0.05)。与Vector组比较,Vector+HLY78组SW480细胞增殖活性显著升高(P<0.05),迁移与侵袭细胞数量显著增多(P<0.05),细胞中β-catenin、c-Myc和MMP-9蛋白表达水平均显著升高(P<0.05)。与Oe-ABCA8组比较,Oe-ABCA8+HLY78组SW480细胞增殖活性显著升高(P<0.05),迁移与侵袭细胞数量显著增多(P<0.05),细胞中β-catenin、c-Myc和MMP-9蛋白表达水平均显著升高(P<0.05)。结论:ABCA8在CRC组织和细胞系中低表达,过表达ABCA8可抑制CRC细胞系SW480的增殖、迁移与侵袭,其作用机制可能与抑制Wnt/β-catenin信号传导有关。
Objective:To study the effect of ATP binding cassette subfamily A member 8(ABCA8)on the proliferation,migration,and invasion of colorectal cancer(CRC)cells,and to clarify its mechanism.Methods:The expression of ABCA8 in CRC tissues was analyzed using bioinformatics,and real-time fluorescence quantitative PCR(RT-qPCR)was used to detect the expression level of ABCA8 mRNA in clinical CRC cancer tissues and various CRC cell lines.SW480 cells were infected with the overexpression of ABCA8 lentivirus,and then divided into Blank group,Vector group,ABCA8 overexpression(Oe-ABCA8)group,Vector+HLY78 group,and Oe-ABCA8+HLY78 group.The intervention concentration of the Wnt/β-catenin signaling pathway agonist HLY78 was 10μmol/L.CCK-8 assay was used to detect cell proliferation activity in each group.Transwell assay was used to detect the number of cell migration and invasion in each group.Western blot method was uesd to detect the protein expression levels of ABCA8,β-catenin,c-Myc,and metalloproteinase-9(MMP-9).Results:ABCA8 showed low expression levels in CRC cancer tissue and CRC cell lines SW480,SW620,HCT116,and HT-29.Compared with the Blank group,the proliferation activity of SW480 cells in the oe-ABCA8 group was significantly reduced(P<0.05),the number of migrating and invading cells was significantly reduced(P<0.05),and the expression of ABCA8 in cells was significantly increased(P<0.05).The protein expression levels ofβ-catenin,c-Myc,and MMP-9 were significantly reduced(P<0.05).However,there was no significant difference in all indicators in the Vector group.Compared with the Vector group,the proliferation activity of SW480 cells in the Vector+HLY78 group were significantly increased(P<0.05),and the number of migrating and invading cells was significantly increased(P<0.05).The protein expression levels of β-catenin,c-Myc,and MMP-9 were significantly increased(P<0.05).Compared with the Oe-ABCA8 group,the proliferation activity of SW480 cells in the Oe-ABCA8+HLY78 group were significantly increased(P<0.05),and the number of migrating and invading cells was significantly increased(P<0.05).The protein expression levels of β-catenin,c-Myc,and MMP-9 were significantly increased(P<0.05).Conclusion:ABCA8 is low expressed in CRC tissues and cell lines,and overexpression of ABCA8 can inhibit the proliferation,migration,and invasion of CRC cell line SW480.Its mechanism may be related to the inhibition of Wnt/β-catenin signaling.
作者
彭肃
阳乐彬
单汉国
PENG Su;YANG Lebin;SHAN Hanguo(Department of Gastrointestinal Surgery,the Second Affiliated Hospital,Hengyang Medical School,University of South China,Hunan Hengyang 421001,China)
出处
《现代肿瘤医学》
CAS
2024年第10期1781-1786,共6页
Journal of Modern Oncology
基金
湖南省卫生健康委卫生科研课题(编号:D202304017061)。
作者简介
彭肃(1986-),男,湖南岳阳人,主治医师,主要从事结直肠肿瘤方面研究。E-mail:pengsu1234@126.com。
