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Archive > Volume 25 Issue 11 > 2018,25(11):1171-1175. DOI:10.3872/j.issn.1007-385X.2018.11.015 Prev Next
Clinical Research
Expression ofHOXA10 gene and its biological effects in endometrial carcinoma
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Abstract:
Objective: To study the expression of HOXA10 gene in endometrial carcinoma and its effect on the apoptosis, migration and invasion of Ishikawa cells. Methods: Twenty-one cases of endometrial carcinoma tissue samples and 25 cases of normal endometrial tissue samples from patients treated at the Department of Obstetrics and Gynecology, Nanjing Drum Tower Hospital from 2012 to 2013 were collected for this study. The mRNA and protein expressions of HOXA10 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, 20 MOI), and infected by adenovirus-flag-lacz (20 MOI) as control; And the cell apoptosis was tested by Flow Cytometry. Ishikawa cells were transfected with 50 nmol/L si-HOXA10 plasmids and 50 nmol/L si-NC plasmids, as down-regulation group and down-regulation control group, respectively. Ishikawa cells were infected with 20 MOI adenovirus-flag-HOXA10 and 20 MOI adenovirus-flag-lacz, as up-regulation group and up-regulation control group, respectively. The ability of migration and invasion was detected by transwell assay. Results: The results of qRT-PCR and Western blotting showed that the expressions of HOXA10 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 HOXA10 gene in Ishikawa cell line, the cell apoptosis rate in ad-flag-HOXA10 groups (5, 10, 20 MOI) 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-HOXA10 group [(248±25) vs (135±15), P<0.01] but markedly down-regulated in ad-flag-HOXA10 group [(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] but markedly down-regulated in ad-flag-HOXA10 group [(34±8) vs (60±4), P<0.01]. Conclusions: Both mRNA and protein expressions of HOXA10 were down-regulated in endometrial carcinoma samples than in normal endometrium. Up-regulation of HOXA10 gene in Ishikawa cell line can promote cell apoptosis and inhibit cell migration and invasion.
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Project Supported:
Project supported by the National Natural Science Foundation of Jiangsu Province（No. BK20151096）
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