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Archive > Volume 21 Issue 3 > 2014,21(3):303-308. DOI:10.3872/j.issn.1007-385X.2014.03.012 Prev Next
Basic Research
The effects and mechanisms of miR-125b overexpression on the proliferation, cycle progression and apoptosis of endometrial carcinoma cell line HEC-1B
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Abstract:
To investigate the effects and underlying mechanisms of miR-125b overexpression on proliferation and apoptosis of endometrial carcinoma (EC) HEC-1B cells in vitro. Methods: Paired EC tissue and adjacent tissue specimens were collected from 30 patients with EC who were treated in the Department of Gynecology and Obstetrics, Sichuan Women’s and Children's Hospital between November 2012 and November 2013. Levels of miR-125b mRNA in these specimens were assessed by real-time PCR. To elucidate the effect and mechanisms of miR-125b overexpression on EC cell death/survival, HEC-1B cells were transfected with an expression vector containing a mimic fragment of miR-125b and a control vector with a sequenced-scrambled fragment, respectively, using lipofectamine were, and cell proliferation, cell cycle progression/apoptosis, and protein levels of PIK3CD, p-AKT and total Akt were assessed by cell counting kit-8 (CCK-8) assays, flow cytometery and Western blotting analysis, respectively, in the transfectants and wild type HEC-1B cells. Results: In all 30 paired specimens, miR-125b mRNA abundance was significantly lower in the endometrial cancer tissue than in the adjacent normal tissue (P<0.05). Transfection of HEC-1B cells with the miR-125b vector resulted in an increase in miR-125b mRNA by more than 820 times. The proliferation index was 0.53±0.06 in HEC-1B cells overexpressing miR-125b, significantly higher (P<0.05) than that in HEC-1B cells overexpressing the sequence-scrambled fragment (0.82±0.07) and wild-type HEC-1B cells (0.89±0.08). The rate of apoptosis with G1 phase arrest was significantly higher in miR-125b-transfected HEC-1B cells as compared with control vector-transfected and wild-type HEC-1B cells ([21.5±3.2]% vs [14.2±2.3]% and [13.5±2.1]%, P<0.01). While no significant difference was observed in total Akt protein content among the three groups of HEC-1B cells (P>0.05), protein contents of PIK3CD and p-AKT were significantly reduced in HEC-1B cells transfected with miR-125b compared with cells transfeced with the control vector and untreated HEC-1B cells (P<0.01). Conclusion: The expression of miR-125b is significantly decreased in the EC tissue. Overexpression of miR-125b may suppress cell proliferation and cell cycle progression and induce cell apoptosis , possibly through suppressing the PI3K/AKT signaling pathway, in HEC-B cells in vitro.
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