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Human amniotic epithelial cells inhibit granulosa cell apoptosis induced by chemotherapy and restore the fertility.

Zhang Q, Xu M, Yao X, Li T, Wang Q, Lai D - Stem Cell Res Ther (2015)

Bottom Line: Premature ovarian failure and insufficiency are significant long-term side-effects of chemotherapy for female cancer patients.Additionally, the ovarian function and fertility of mice were assessed via counts of follicles and mating experiments at 4 weeks after hAEC transplantation. hAECs significantly inhibited tumor necrosis factor-alpha-mediated granulosa cell apoptosis induced by chemotherapeutics and reduced the inflammatory reaction in ovaries at 7 days after transplantation.These results suggest a potential molecular mechanism for the effective therapy of hAEC transplantation in chemotherapy-induced premature ovarian failure and insufficiency.

View Article: PubMed Central - PubMed

Affiliation: International Peace Maternity and Child Health Hospital, School of Medicine, Shanghai Jiao Tong University, 145, Guang-Yuan Road, Shanghai, 200030, People's Republic of China. 10111010022@fudan.edu.cn.

ABSTRACT

Introduction: Premature ovarian failure and insufficiency are significant long-term side-effects of chemotherapy for female cancer patients. Recently, stem cell transplantation has been identified as a promising treatment for premature ovarian failure and insufficiency. We have previously demonstrated that human amniotic epithelial cells (hAECs) migrate into injured tissue and promote the recovery of ovarian function in chemoablated mice. However, the molecular mechanism guiding this process remains unclear.

Methods: To further investigate the effect of hAECs on chemotherapy-induced apoptosis, cultured primary hAECs were injected intravenously into mice treated with cyclophosphamide and busulphan. Apoptosis of granulosa cells was observed by TUNEL staining, and apoptosis-related gene expression was performed on ovarian tissue by real-time PCR and Western blot 7 days after hAEC transplantation. Additionally, the ovarian function and fertility of mice were assessed via counts of follicles and mating experiments at 4 weeks after hAEC transplantation.

Results: hAECs significantly inhibited tumor necrosis factor-alpha-mediated granulosa cell apoptosis induced by chemotherapeutics and reduced the inflammatory reaction in ovaries at 7 days after transplantation. In addition, 4 weeks after transplantation, hAECs promoted the development of follicles and increased the number of cumulus oocyte complexes in chemoablated mice. Furthermore, hAECs improved ovarian mass and increased the number of follicles compared to those of the chemoablated group, and hAEC transplantation partially rescued the fertility of chemoablated mice.

Conclusions: hAEC transplantation promotes ovarian function by inhibiting tumor necrosis factor-alpha-mediated cell apoptosis and reducing inflammation in chemotherapy-induced premature ovarian failure. These results suggest a potential molecular mechanism for the effective therapy of hAEC transplantation in chemotherapy-induced premature ovarian failure and insufficiency.

No MeSH data available.


Related in: MedlinePlus

Chemotherapy-induced apoptosis of granulosa cells within developing follicle. A TUNEL-positive cells were observed in the ovary tissue at 7 days after chemotherapy. The red stain indicates TUNEL-positive granulosa cells. The blue DAPI stain indicates the cell nucleus. B Relative expression of Bcl2, Bax and TNF-α mRNA in ovarian tissue at 3, 7 and 10 days after chemotherapy. Data represent means ± SEM; *p < 0.05 versus Sham. Scale bar = 200 μm (a, b, c and d), 100 μm (e and f). Sham sham group (n = 6), Cy-3days chemoablated 3-day group (n = 6), Cy-7days chemoablated 7-day group (n = 6), Cy-10days chemoablated 10-day group (n = 6)
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Fig2: Chemotherapy-induced apoptosis of granulosa cells within developing follicle. A TUNEL-positive cells were observed in the ovary tissue at 7 days after chemotherapy. The red stain indicates TUNEL-positive granulosa cells. The blue DAPI stain indicates the cell nucleus. B Relative expression of Bcl2, Bax and TNF-α mRNA in ovarian tissue at 3, 7 and 10 days after chemotherapy. Data represent means ± SEM; *p < 0.05 versus Sham. Scale bar = 200 μm (a, b, c and d), 100 μm (e and f). Sham sham group (n = 6), Cy-3days chemoablated 3-day group (n = 6), Cy-7days chemoablated 7-day group (n = 6), Cy-10days chemoablated 10-day group (n = 6)

Mentions: To interrogate the mechanism contributing to chemotherapy-induced loss of follicles, we investigated the effects of chemoablation on cell apoptosis in ovarian tissue. TUNEL-positive cells were observed in the ovarian section of mice at 7 days after chemotherapy, suggesting apoptosis was largely restricted to the GC layer of secondary follicle, in proximity to the oocytes (Fig. 2A). To this effect, we observed significant downregulation of the antiapoptotic gene Bcl2 in ovaries of chemoablated mice (Fig. 2B, p < 0.05). Studies have demonstrated that ovaries can express tumor necrosis factor (TNF) receptors, and is sensitive to the TNF-α-mediated cell apoptosis pathway [22]. In order to further investigate whether TNF-α participates in chemotherapy-induced apoptosis, we also evaluated the expression of TNF-α in the injured ovaries. TNF-α mRNA was significantly increased in the ovaries of chemoablated mice as compared to controls (Fig. 2B, p < 0.05). These results showed that chemotherapy drugs induce GC apoptosis, and increase inflammation within ovarian tissue.Fig. 2


Human amniotic epithelial cells inhibit granulosa cell apoptosis induced by chemotherapy and restore the fertility.

Zhang Q, Xu M, Yao X, Li T, Wang Q, Lai D - Stem Cell Res Ther (2015)

Chemotherapy-induced apoptosis of granulosa cells within developing follicle. A TUNEL-positive cells were observed in the ovary tissue at 7 days after chemotherapy. The red stain indicates TUNEL-positive granulosa cells. The blue DAPI stain indicates the cell nucleus. B Relative expression of Bcl2, Bax and TNF-α mRNA in ovarian tissue at 3, 7 and 10 days after chemotherapy. Data represent means ± SEM; *p < 0.05 versus Sham. Scale bar = 200 μm (a, b, c and d), 100 μm (e and f). Sham sham group (n = 6), Cy-3days chemoablated 3-day group (n = 6), Cy-7days chemoablated 7-day group (n = 6), Cy-10days chemoablated 10-day group (n = 6)
© Copyright Policy - OpenAccess
Related In: Results  -  Collection

License 1 - License 2
Show All Figures
getmorefigures.php?uid=PMC4549019&req=5

Fig2: Chemotherapy-induced apoptosis of granulosa cells within developing follicle. A TUNEL-positive cells were observed in the ovary tissue at 7 days after chemotherapy. The red stain indicates TUNEL-positive granulosa cells. The blue DAPI stain indicates the cell nucleus. B Relative expression of Bcl2, Bax and TNF-α mRNA in ovarian tissue at 3, 7 and 10 days after chemotherapy. Data represent means ± SEM; *p < 0.05 versus Sham. Scale bar = 200 μm (a, b, c and d), 100 μm (e and f). Sham sham group (n = 6), Cy-3days chemoablated 3-day group (n = 6), Cy-7days chemoablated 7-day group (n = 6), Cy-10days chemoablated 10-day group (n = 6)
Mentions: To interrogate the mechanism contributing to chemotherapy-induced loss of follicles, we investigated the effects of chemoablation on cell apoptosis in ovarian tissue. TUNEL-positive cells were observed in the ovarian section of mice at 7 days after chemotherapy, suggesting apoptosis was largely restricted to the GC layer of secondary follicle, in proximity to the oocytes (Fig. 2A). To this effect, we observed significant downregulation of the antiapoptotic gene Bcl2 in ovaries of chemoablated mice (Fig. 2B, p < 0.05). Studies have demonstrated that ovaries can express tumor necrosis factor (TNF) receptors, and is sensitive to the TNF-α-mediated cell apoptosis pathway [22]. In order to further investigate whether TNF-α participates in chemotherapy-induced apoptosis, we also evaluated the expression of TNF-α in the injured ovaries. TNF-α mRNA was significantly increased in the ovaries of chemoablated mice as compared to controls (Fig. 2B, p < 0.05). These results showed that chemotherapy drugs induce GC apoptosis, and increase inflammation within ovarian tissue.Fig. 2

Bottom Line: Premature ovarian failure and insufficiency are significant long-term side-effects of chemotherapy for female cancer patients.Additionally, the ovarian function and fertility of mice were assessed via counts of follicles and mating experiments at 4 weeks after hAEC transplantation. hAECs significantly inhibited tumor necrosis factor-alpha-mediated granulosa cell apoptosis induced by chemotherapeutics and reduced the inflammatory reaction in ovaries at 7 days after transplantation.These results suggest a potential molecular mechanism for the effective therapy of hAEC transplantation in chemotherapy-induced premature ovarian failure and insufficiency.

View Article: PubMed Central - PubMed

Affiliation: International Peace Maternity and Child Health Hospital, School of Medicine, Shanghai Jiao Tong University, 145, Guang-Yuan Road, Shanghai, 200030, People's Republic of China. 10111010022@fudan.edu.cn.

ABSTRACT

Introduction: Premature ovarian failure and insufficiency are significant long-term side-effects of chemotherapy for female cancer patients. Recently, stem cell transplantation has been identified as a promising treatment for premature ovarian failure and insufficiency. We have previously demonstrated that human amniotic epithelial cells (hAECs) migrate into injured tissue and promote the recovery of ovarian function in chemoablated mice. However, the molecular mechanism guiding this process remains unclear.

Methods: To further investigate the effect of hAECs on chemotherapy-induced apoptosis, cultured primary hAECs were injected intravenously into mice treated with cyclophosphamide and busulphan. Apoptosis of granulosa cells was observed by TUNEL staining, and apoptosis-related gene expression was performed on ovarian tissue by real-time PCR and Western blot 7 days after hAEC transplantation. Additionally, the ovarian function and fertility of mice were assessed via counts of follicles and mating experiments at 4 weeks after hAEC transplantation.

Results: hAECs significantly inhibited tumor necrosis factor-alpha-mediated granulosa cell apoptosis induced by chemotherapeutics and reduced the inflammatory reaction in ovaries at 7 days after transplantation. In addition, 4 weeks after transplantation, hAECs promoted the development of follicles and increased the number of cumulus oocyte complexes in chemoablated mice. Furthermore, hAECs improved ovarian mass and increased the number of follicles compared to those of the chemoablated group, and hAEC transplantation partially rescued the fertility of chemoablated mice.

Conclusions: hAEC transplantation promotes ovarian function by inhibiting tumor necrosis factor-alpha-mediated cell apoptosis and reducing inflammation in chemotherapy-induced premature ovarian failure. These results suggest a potential molecular mechanism for the effective therapy of hAEC transplantation in chemotherapy-induced premature ovarian failure and insufficiency.

No MeSH data available.


Related in: MedlinePlus