Mitigation of premature ovarian failure by over-expression of lentivirus vector-mediated Wilms tumor-suppressor gene

Juan Wang,Bin Jiang,Le Bo,Wendan Xu,Xuekai Li,Caiping Mao

doi:10.4314/tjpr.v17i9.9

Abstract

Purpose: To investigate the effect of WT1(Wilms tumor-suppressor gene) overexpression on premature ovarian failure (POF)-mediated ovarian dysfunction. Methods: Three mice groups were used: control group (untreated mice), POF group (mice sterilized by intravenous injection of cyclophosphamide and busulfan), and POF-LV(lentiviral vector)􀀀GFP(green fluorescent protein)􀀀WT1 group (POF mice given intra-ovarian microinjection of LV􀀀GFP􀀀WT1, a WT1􀀀overexpressing lentiviral vector, one week after sterilization). Real time-PCR was employed to analyze in vitro WT1 overexpression levels. Overall ovarian function was measured by hormonal assay, H & E staining, and immuno-histochemical techniques. Results: Overexpression of WT1 in mice models of POF alleviated ovarian granulosa cell (GC) damage, increased ovary weight, and significantly increased follicular number (p < 0.05). Radioimmunoassays revealed reduction in plasma estradiol (E2) and follicle-stimulating hormone (FSH, p < 0.05). However, results from immune-histochemical assays showed reduced Bax expression levels, and increased expression of Bcl-2 in WTI-overexpression mice, relative to POF mice. Conclusion: Overexpression of WT1 may stimulate repair of ovarian tissue while improving endocrine function by inhibiting ovarian cell apoptosis signaling pathway in POF mice. Keywords: Premature ovarian failure, Wilms tumor suppressor gene, Chemotherapy, Apoptosis, Estradiol

Highlights

Chemotherapy-induced premature ovarian failure (POF) and infertility are areas of immense interest to clinicians
Chemotherapy-induced POF and infertility are areas of immense interest to clinicians. This is due to rising incidence of malignant tumors and chemotherapy-induced POF, as well as increasing population of relatively young patients with malignant ovarian tumors [1,2]
A 90 % lentiviral infection efficiency was successfully effected in granulosa cell (GC) by LV-GFP-WT1 treatment, as confirmed by quantification of GFP expression 72 h after transfection (Figure 2 A)

Summary

Introduction

Chemotherapy-induced POF and infertility are areas of immense interest to clinicians. This is due to rising incidence of malignant tumors and chemotherapy-induced POF, as well as increasing population of relatively young patients with malignant ovarian tumors [1,2]. Apart from POF and infertility, chemotherapy is accompanied by complications such as anorexia, amenorrhea, cardiovascular diseases and osteoporosis, all of which severely impact the life quality of young cancer survivors [3]. Fertilityenhancing treatments are available for POF patients. These include in vitro fertilization and hormonal therapy [4]. There are limited options for patients with chemotherapyinduced POF.

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Results

Conclusion