Folicle-Stimulating Hormone Promotes The Migration Of SK-OV-3 Ovarian Cancer Cells Via Upregulation Of Cystathionine-β-synthase

BackgroudOvarian cancer is a common gynecologic cancer.Ovarian cancer is one of the three most common maligancies of female genitalia Due to the fact that the ovaries are located in the deep pelvic cavity,early lesions are not easy to be found,and there is a lack of effective treatments for the advanced ovarian cancer.Therefore,the mortality rate of ovarian cancer ranks first in gynecologic cancer,and it has become the main cancer that seriously threatens the life and health of the women.Folicle-stimilating hormone(FSH)is a type of glycoprotein hormone that secreted by the anterior pituitary basophilic granulocyte.Folicle-stimilating hormone receptor(FSHR)express in a variety of cells.In recent years,more and more studies show that FSH not only plays a role in the classic reproductive regulation but also plays a more important role beside the gonads.And FSH takes participation in the occurrence and development of varieties of disease.There are large number of researches have identified that FSHR express in the epithelial cells and peripheral vascular endothelial cells of ovarian cancer、prostate cancer、breast cancer and other cancers,which may regulate the Proliferation、migration and invasion of the cancer.But the mechanism of FSHR’s role in tumorigenesis and its use as a new therapeutic target remains to be further studies.Hydrogen sulfide(H2S)has been recognized as the third kind of gas signal molecule following the nitric oxide(NO)and carbon monoxide(CO).H2S is synthesised by cystathionine synthase(CBS)、cystathionine lyase(CSE)and 3-thiopyruvate thiotransferase(3-MST).CBS in primary serous ovarian cancer is very common CBS is a important part in maintaining the REDOX balance and metabolism in the ovarian cancer cells,so as to promote the growth and metastasis of ovarian cancer.It is still uncleare whether FSH can regulate the activity of CBS protein thus regulate the release of H2S.The specific molecular mechanism of FSH affeccting CBS protein also has not yet been elucidating.Even more,how does CBS plays in the process of FSH affecting ovarian cancer cells’ migration has not been clarified.Based on this,our research mainly studied the molecular mechanism of FSH regulating CBS in ovarian cancer cells and its affect on ovarian cancer cells’ metastasis.ObjectiveTo explore the molecular mechanism of Folicle-stimilating hormone(FSH)regulating CBS,and to clarify the effect of FSH through CBS/H2S on ovarian cancer cells’ metastasis.Methods and ResultSubcultured the SK-OV-3 ovarian cancer cells after we bought it from the company,we used free radical electrode to detected the level of the release of H2S.The results showed that FSH could enhance the release H2S after we treated the SK-OV-3 ovarian cancer cells with FSH(75mIU/mL)for different time or different concentrations of FSH for 24h.Western blotting was used to detected the effect of FSH on CBS,CSE and 3-MST protein levels in SK-OV-3 ovarian cancer cells.It showed that FSH could upregulate CBS protein level,but has no significant effects on CSE and 3-MST protein level.After that,we found that FSH could upregulate CBS mRNA expression which is detected by Quantiative Real-time PCR.Next,we had western blotting,cell migration and cell invasion assay.At this step,we pretreated the SK-OV-3 ovarian cancercells with H2S donor NaSH and CBS inhabitor AOAA for 1h respectively,then treated the SK-OV-3 cancer cells with FSH for 24h.The results proved that compared with the CON group,the NaSH group and AOAA group down-regulated the level of CBS protein,and inhabited the migration of the SK-OV-3 ovarian cancer cells.At the same tine,we transfected CBS siRNA into the SK-OV-3 ovarian cancer cells.we got the results that compared with CON group,CBS si RNA groups’ migration and invasion were inhabited Then we had another treatments for the SK-OV-3 ovarian cancer cells:pretreated the SK-OV-3 ovarian cancercells with SP1 inhabitor MTA,CREB inhabitor 666-15 and NF-κB inhabitor SC75741 for 1h respectively,then treated the SK-OV-3 cancer cells with FSH for 24h.The results indicated that compared with CON group,the level of CBS protein are down-regulated in the MTA group and CREB group;the level of CBS protein are up-regulated in the NF-κB group more obviously;the MTA groups’ migration are inhabited.After transfected CREB siRNA,we got that in the CREB siRNA group,the effects of FSH was inhabited and inhabited the SK-OV-3 ovarian cancer cells migration.Conclusion1.FSH upregulated CBS protein level and mRNA expression,and enhanced H2S release.,which promoted the SK-OV-3 ovarian cancer cells metastasis.2.FSH may mediated the CBS protein expression via SP1 or CREB signaling pathway.