HOXA10基因在子宫内膜癌组织中的表达及其生物学作用

Expression of HOXA10 gene and its biological effects in endometrial carcinoma

目的:探讨同源框A10(HOXA10)基因在子宫内膜癌组织中的表达及其对子宫内膜癌Ishikawa细胞株凋亡、迁移及侵袭的影响。方法:收集2012年至2013年在鼓楼医院妇产科子宫内膜癌组织标本21例、正常增殖期子宫内膜组织标本25例,采用实时荧光定量PCR(qRT-PCR)及Western blotting方法检测HOXA10在子宫内膜癌组织及正常子宫内膜组织中的表达。将感染复数为5、10、20 MOI的ad-flag-HOXA10腺病毒质粒及20 MOI的ad-flag-lacz腺病毒质粒(对照组)感染人子宫内膜癌Ishikawa细胞株,流式细胞术检测各组细胞凋亡情况。将50 nmol/L的si-HOXA10及si-NC质粒转染Ishikawa细胞株,分别为下调组及下调对照组;将20 MOI的ad-flag-HOXA10及20 MOI的ad-flag-lacz腺病毒质粒感染Ishikawa细胞株,分别为上调组及上调对照组;Transwell小室检测各组细胞迁移及侵袭能力。结果:在子宫内膜癌组织中HOXA10 mRNA表达量比正常子宫内膜组织中显著降低([0.56±0.14)vs (1.36±0.33),P<0.01],其蛋白表达量同样显著降低([1.01±0.25)vs (2.10±0.71),P<0.01]。上调HOXA10后5、10、20 MOI组细胞的凋亡率明显升高,且大多数处于早期凋亡([50.92±8.79)%、(55.17±4.07)%、(76.10±3.65)%vs (7.74±0.15)%,均P<0.01]。下调HOXA10表达后迁移细胞数显著增加([248±25)vs (135±15)个,P<0.01],上调HOXA10表达后迁移细胞数显著减少([50±6)vs (100±13)个,P<0.01];下调HOXA10表达后侵袭细胞数显著增多([131±18)vs (66±9)个,P<0.01],上调HOXA10表达后侵袭细胞数显著减少([34±8)vs(60±4)个,P<0.01]。结论:HOXA10基因在子宫内膜癌内膜中表达低于正常内膜,子宫内膜癌Ishikawa细胞株中上调HOXA10基因表达能促进细胞凋亡并抑制其迁移和侵袭能力。

Objective:To study the expression of HOXAIO gene in endometrial carcinoma and its effect on the apoptosis,migration and invasion of Ishikawa cells.Methods:Twenty-one cases of endometrial earcinoma tissue samples and 25cases of normal endometri- al tissue samples from patients treated at the Department of Obstetrics and Gynecology,Nanjing Drum Tower Hospital from 2012to 2013were collected for this study.The mRNA and protein expressions of HOXAIO in endometrial carcinoma and normal endometrial tissues were separately tested by Realtime-qPCR (qRT-PCR)and Western blotting.Ishikawa cells were infected with adenovirus-flag- HOXA10 at different multiplicity (5,10,20MOI),and infected by adenovirus-flag-lacz (20MOI)as control;And the cell apoptosis was tested by Flow Cytometry.Ishikawa cells were transfected with 50nmol/L si-HOXA10 plasmids and 50 nmol/L si-NC plasrnids,as down-regulation group and down-regulation control group,respectively.Ishikawa cells were infected with 20MOI adenovirus-flag- HOXA10 and 20MOI adenovirus-flag-lacz,as up-regulation group and up-regulation control group,respectively.The ability of migra- tion and invasion was detected by transwell assay.Results:The results of qRT-PCR and Western blotting showed that the expressions of HOXAIO mRNA and protein in endometrial carcinoma samples were both significantly lower than normal samples [mRNA:(0.56± 0.14)vs (1.36±0.33),P<0.01;protein:(1.01±0.25)vs (2.10±0.71),P<0.001].After the up-regulation of HOXAIO gene in Ishikawa cell line,the cell apoptosis rate in ad-flag-HOXA10groups (5,10,20MOI)was significantly raised,and most of which was in the early apoptosis [(50.92±8.79)%,(55.17±4.07)%,(76.10±3.65)%vs (7.74±0.15)%,all P<0.01].The number of migrated cells was markedly up-regulated in si-HOXA10group [(248±25)vs (135±15),P<0.01]but markedly down-regulated in ad-flag-HOXA10group [(50±6)vs (100±13),P<0.01].The number of invasive cells was markedly up-regulated in si-HOXA10 group [(131±18)vs (66±9),P<0.01]butmarkedly down-regulated in ad-flag-HOXA10 group [(34±8)vs (60±4),P<0.01].Conclusions:Both mRNA and protein expressions of HOXAlO were down-regulated in endometrial carcinoma samples than in normal endometrium.Up-regulation ofHOXAIO gene in Ishi- kawa cell line can promote cell apoptosis and inhibit cell migration and invasion.