ACTL8表达与子宫内膜癌的发生及细胞侵袭、迁移能力的关系

Relationship between the expression of ACTL8 and the occurrence of endometrial carcinoma and the ability of cell invasion and migration

目的探讨子宫内膜癌组织中肌动蛋白样蛋白8(ACTL8)表达与肿瘤发生及癌细胞侵袭、迁移能力的关系。方法回顾性选取2020年3月至2021年3月恩施土家族苗族自治州中心医院妇儿医院病理科收集的68例子宫内膜癌组织(病例组),并选取45例正常子宫内膜作为对照组。采用蛋白质印迹法检测两组标本中的ACTL8蛋白。选取人子宫内膜癌KLE细胞系,以携带shRNA特异序列的载体及空载质粒分别KLE细胞系,形成敲出ACTL8基因的sh ACTL8-1组、sh ACTL8-2组及以空载质粒转染KLE细胞系的对照组。采用荧光定量PCR检测3组细胞中ACTL8 RNA表达,采用蛋白质印迹法检测3组细胞ACTL8蛋白表达,采用Transwell迁移和侵袭实验分析KLE细胞系培养后的侵袭和迁移能力差异。结果病例组标本中的ACTL8蛋白的相对表达强度为0.983±0.211,对照组标本中的ACTL8蛋白的相对表达强度为0.377±0.089,子宫内膜癌组织中的ACTL8蛋白相对表达强度显著高于对照组,差异有统计学意义(P<0.05)。在年龄≥55岁、子宫内膜癌分级G3级、FIGO分期(Ⅲ期+Ⅳ期)的子宫内膜癌组织中的ACTL8蛋白的相对表达强度高于<55岁、子宫内膜癌分级(G1+G2)、FIGO分期(Ⅰ期+Ⅱ期)的患者,差异均有统计学意义(P<0.05)。对照组的ACTL8 RNA、ACTL8蛋白表达显著高于sh ACTL8-1组、sh ACTL8-2组,差异有统计学意义(P<0.05);sh ACTL8-1组、的ACTL8 RNA、ACTL8蛋白表达显著高于sh ACTL8-2组,差异有统计学意义(P<0.05)。sh ACTL8组的侵袭、迁移细胞数目均低于对照组,差异均有统计学意义(P<0.05)。结论子宫内膜癌组织中ACTL8蛋白表达上调并且与肿瘤发生发展关系密切;ACTL8基因表达与肿瘤细胞侵袭及迁移能力有关。

Objective To investigate the relationship between the expression of actin-like protein 8(ACTL8)in endometrial cancer tissue and tumorigenesis and the ability of cancer cell invasion and migration.Methods A total of 68 cases of endometrial cancer tissues(case group)collected from the Department of Pathology,Women and Children's Hospital of Enshi Tujia and Miao Autonomous Prefecture Central Hospital from March 2020 to March 2021 were retrospectively selected,and 45 cases of normal endometrium were selected as the control group.The ACTL8 protein in the two groups of specimens was detected by Western blotting.The human endometrial cancer KLE cell line was selected,and the vector carrying the shRNA-specific sequence and the empty vector plasmid were used to separate the KLE cell lines to form the sh ACTL8-1 group that knocked out the ACTL8 gene.In the sh ACTL8-2 group and the control group transfected with KLE cell line with the empty plasmid,the expression of ACTL8 RNA in the cells of the three groups was detected by fluorescence quantitative PCR,the protein expression of ACTL8 in the cells of the three groups was detected by Western blotting method,and differences in invasion and migration abilities of KLE cell lines after culture were analyzed using Transwell migration and invasion assays.Results The relative expression intensity of ACTL8 protein in the specimens of the case group was 0.983±0.211,the relative expression intensity of ACTL8 protein in the specimens of the control group was 0.377±0.089,and the relative expression intensity of ACTL8 protein in endometrial cancer tissues was significantly higher than that in the control group,the difference was statistically significant(P<0.05).The relative expression intensity of ACTL8 protein in endometrial cancer tissues aged≥55 years,endometrial cancer grade G3,and FIGO stage(Ⅲ+Ⅳ)was higher than that of aged<55 years old,endometrial cancer grade(G1+G2)and FIGO staging(stage Ⅰ+stage Ⅱ),the differences were statistically significant(P<0.05).The expressions of ACTL8 RNA and ACTL8 protein in the control group were significantly higher than those in the sh ACTL8-1 group and sh ACTL8-2 group,the differences were statistically significant(P<0.05);the expressions of ACTL8 RNA and ACTL8 protein in the sh ACTL8-1 group and ACTL8-1 group were higher than those in the sh ACTL8-2 group,the differences were statistically significant(P<0.05);the number of invasive and migrating cells in the sh ACTL8 group was lower than that in the control group,and the difference was statistically significant(P<0.05).Conclusion The expression of ACTL8 protein in endometrial cancer tissues is up-regulated and is closely related to tumorigenesis and development;the expression of ACTL8 gene is related to tumor cell invasion and migration ability.