| Objective: The occurrence and development of breast cancer is affected by many factors,and its incidence is increasing year by year.It becomes the highest incidence of malignant tumors in the world.The treatment of breast cancer is still more traditional and single.Molecular targeting therapy has become a hot spot in clinical research,but the biomarkers and molecular targets of breast cancer are still very limited.In order to screen valuable breast cancer related factors,a new gene RRS1(Regulator of Ribosome Synthesis 1),is successfully screened for breast cancer sample information through high throughput differential expression gene screening and bioinformatics analysis.To explore the correlation between RRS1 gene and the occurrence and development of breast cancer by detecting the expression difference of RRS1 gene in human breast normal epithelial cells HMEC and breast cancer cells MCF-7.The effect of RRS1 on the proliferation,apoptosis and migration of human breast cancer cell MCF-7 was analyzed by the inhibition of the expression of RRS1 by the RNA interference mediated by lentivirus.Methods: Western blot assey were used to detect the RRS1 gene expression differences between MCF-7 and normal mammary epithelial HMEC cells.Lentiviral-mediated RNA interference system with green fluorescent protein were constructed with RRS1-sh RNA virus vector and negative control virus vector,and then infected MCF-7 cells,respectively.After that,fluorescence microscopy were used to determine the infection efficiency;RRS1 expression levels were detected by Real-time PCR and Western blot.MTT assey were sued to detect the proliferation activity for six consecutive days;PI FCM was used to detect the distribution of cell cycle;morphology and molecular level of apoptosis of the two groups were examined by DAPI fluorescent staining,Annexin V-APC single dye FCM,and Caspase 3/7 assey;cell migration ability were detected by transwell assey.Results: RRS1 protein expression levels of MCF-7 were apparently higher than HMEC(P<0.0001).Compared with the control sh Ctrl group,both the m RNA and protein level of sh RRS1 were decreased(P<0.05),this shows that the sh RNA-RRS1 lentivirusvector can effectively inhibit the expression of RRS1 gene.Cells proliferation significantly reduced(P<0.001),proportion of G1 phase increased(P<0.05).The cells after knocking down the expression of RRS1 had typical morphological changes of apoptosis.The fluorescence of the nucleus in the sh Ctrl control group was uniform bright blue circular or oval,and dense dense particles were found in most of the nuclei and cytoplasm in the sh RRS1 experimental group.Annexin V-APC single staining flow cytometry showed a significant increase in apoptosis rate after subtraction(P<0.001),and Caspase3/7 test showed that the Caspase3/7 activity in sh RRS1 group was significantly higher than that in sh Ctrl group,and the difference was significant(P<0.0001).At the same time,compared with the sh Ctrl group,the ability of cell migration decreased significantly after RRS1 knockdown(P<0.0001).Conclusion: RRS1 gene is highly expressed in breast cancer MCF-7 cell.Knocking down the expression of RRS1 gene can significantly induce the proliferation and apoptosis of breast cancer cell MCF-7,as well as cell cycle G1 arrest;meanwhile,the migration ability of breast cancer cells is significantly reduced.It is indicated that RRS1 is related to the proliferation,apoptosis and cycle process of breast cancer cells.Considering the role of RRS1 in ribosome biosynthesis and other mechanisms,this results provides another significant research direction for molecular targeted therapy of breast cancer. |
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