Sulforaphane Enhances The Radiosensitivity Of Cervical Cancer Cells By Inhibiting DNA Repair Via LATS2-Rad51 Signaling Pathway & Inhibition Of Sulforaphane On The Proliferation Of Cervical Cancer HeLa Cells Via Up-regulating RASD1

Sulforaphane enhances the radiosensitivity of cervical cancer cells by inhibiting DNA repair via LATS2-Rad51 signaling pathwayBackground and purpose: Cervical cancer is the most commonly diagnosed gynecological cancer,and the leading cause of deaths from gynecological malignancies in developing countries,it has been rejuvenated in recent years.Radiotherapy is the main treatment for advanced patients with cervical squamous cell carcinoma,and some patients are not sensitive to Radiotherapy,which remains the biggest obstacle to the treatment of cervical cancer.The sulforaphane induces the cell cycle arrest at G2/M phase,by regulating various cell enzyme systems,and promots the cancer cells apoptosis,it can inhibit tumor angiogenesis and the metastasis and spread of cancer cells too,the underlying mechanism remains elusive.A recent study reported that sulforaphane enhances the tumor cells radiosensitivity,However,the underlying mechanism of sulforaphane in Cervical cancer cells remains largely unknown.This study aimed to investigate the potential mechanism of SFN enhances cervical cancer cells radiosensitivity.Methods: The effect of SFN on the cell viability was assessed by CCK8 method,search for the target(s)for SFN,the m RNA sequencing was conducted to screened out a target of SFN treated with Si Ha cells.Then the target of SFN was confirmed by western blot,q RT-PCR and luciferase assays.Cells were treated with SFN or not,transfected with two sequences of LATS2 overexpression or control prior.Cell apoptosis rates were assessed with flow cytometric analysis.We used western blot to detect the protein expression of cleaved caspase-3 and cleaved-par.Then we transfected cells with LATS2 overexpression prior,control prior,sh LATS2 and sh LATS2 prior plus SFN,exposed to ionizing Radiation,cell proliferation was assessed by clonogenic assays.Immunofluorescence analysis and Western blot to detect the protein expression of γ-H2 AX,MDC1 and Rad51.Results: 1.SFN inhibits the proliferation of cervical cancer cells and treatment increased LATS2 expression in cervical cancer cells.2.LATS2 suppresses the proliferation of cervical cancer Si Ha and C33 A cells,SFN and LATS2 promote cells apoptosis.3.Post-Radiotherapy,SFN promoted the expression of γ-H2 AX through DNA double-strand break,and inhibited the aggregation of MDC1 and Rad51 in the nucleus.4.Post-Radiotherapy,transfected with LATS2 had no significant effect on cell DNA double-strand break,but inhibited the aggregation of MDC1 and Rad51 in the nucleus,and in si LATS2 cells,the aggregation in the nuclear was more than the control group.Conclusion: This experiment demonstrates that SFN inhibits the proliferation of cervical cancer cells.Our data firstly reveals that SFN treatment increased LATS2 expression in cervical cancer cells,and LATS2 inhibits the proliferation and promotes apoptosis.Post-Radiotherapy,LATS2 blocks Rad51 recruitment to the DNA double strand break site.These results provide novel experimental support for SFN regulating the expression of LATS2 enhances the radiosensitivity of cervical cancer cells.Inhibition of sulforaphane on the proliferation of cervical cancer He La cells via up-regulating RASD1Background and purpose: Cervical cancer is one of the three major malignant tumors of gynecology.It has shown a trend of rejuvenation in recent years and is the leading cause of cancer death in women in developing countries.It has been found that sulforaphane(SFN)regulates tumor cell cycle arrest,oxidative stress and apoptosis by different metabolic systems at different stages of the cell,but its anticancer mechanism hasn’t been determined.Studies have shown that RASD1 can induce cell cycle arrest,inhibit cell growth,and inhibit cell migration and invasion in tumor cells.In the previous experiments,we found that SFN can promote the expression of RASD1.Therefore,this study uses cervical cancer He La cells to investigate the function and the potential mechanism of SFN-induced RASD1 on the apoptosis and cycle of cervical cancer cells.Methods: He La cells were treated with different concentrations of SFN for 48 h.Cell proliferation was measured by CCK-8 assay.The m RNA and protein expression of RASD1 in He La cells were detected by real time-PCR and Western blot,respectively.Then He La cells were treated with SFN or not,transfected with RASD1 and Control vector for 48 h,cells apoptosis and cycle were detected by Flow cytometry,Western blot detected the relative protein level.and decreasing the level of cdc2 protein(P<0.05).Results: 1.SFN inhibited the proliferation of He La cells in a concentration-time dependent manner.Furthermore,SFN treatment could induce the expression of RASD1 at the m RNA and protein levels.2.The apoptosis ratio of cells treated with SFN or over-expression RASD1 cells was significantly higher than their control group,and the expression levels of cleaved-caspase3 and cleaved-parp were up-regulated than control group.3.Cells treated with SFN or RASD1 over-expression group,the percentage of cells in S phase less than control group,and more than in G2/M phase clearly.Western blot shows cells with SFN or over expressing RASD1 remarkable increasing the level of p21 proteinConclusion: This experiment demonstrates that SFN inhibits the proliferation of cells.Our data firstly reveals that SFN treatment increased RASD1 expression in cervical cancer cells,and RASD1 promotes apoptosis and cell cycle arrest,and promotes apoptosis.Provide novel experimental support for SFN against cervical cancer,may be related to the increased expression of RASD.