Ginsenoside Rh1 Inhibits The Malignant Biological Behavior Of HER2-positive Breast Cancer Cells Through DDX5

Objective:The aim of this study was to investigate the effects of ginsenoside Rh1 on HER2 positive breast cancer cell line SKBR3 at the cellular levels.1.To investigate the effect of ginsenoside Rh1 on malignant biological behavior and DDX5 expression of breast cancer SKBR3 cells.2.To detect the expression of DDX5 in breast cancer and its relationship with clinical stage,pathological grade.Methods:1.MTS assay was used to detect ginsenoside Rh1(10 、50 、100、500 uMol).The effect on the proliferation of breast cancer SKBR3 cells,the selection of meaningful drug concentration and action time.2.The effect of ginsenoside Rh1 on the migration of breast cancer SKBR3 cells at 24 h or 48 h was observed by scratch test.3.The effect of ginsenoside Rh1 on the invasive ability of breast cancer SKBR3 cells at 24 h was observed by Transwell invasion assay.4.Flow cytometry was used to observe the effect of ginsenoside Rh1 on apoptosis and cell cycle of breast cancer SKBR3 cells at 48 h.5.The effect of ginsenoside Rh1 in 48 h was detected by Western blot(Western-blot)in the expression of DDX5 protein in SKBR3 cells of breast cancer.6.Effect of ginsenoside Rh1 on the expression of DDX5 gene in breast cancer SKBR3 cells after 48 h detection by fluorescent quantitative PCR.7.From December 1st 2014 to 31 December 2017,98 patients with breast cancer were collected and screened from Shenyang military region General Hospital,who were not treated with radiotherapy and chemotherapy,traditional Chinese medicine and other treatments,with detailed pathological data and paraffin samples of cancer tissue.The expression of DDX5 in breast cancer tissues was detected by immunohistochemical method,and the expression of DDX5 in breast cancer tissues was detected by immunohistochemical method,which combined with clinicopathological data,35 cases of breast cancer patients with HER2 overexpression [amplified by HER2 detected by Fish] were treated with HER-negative breast cancer.To observe and analyze the relationship between DDX5 and molecular classification,clinical stage,pathological grade and other clinicopathological features of breast cancer.Results:1.The effect of ginsenoside Rh1 on the growth of HER2 positive breast cancer cells.The results showed that 500 u M ginsenoside Rh1 inhibited the proliferation of HER2 positive human breast cancer cell line(SKBR3)at 24 h or 48 h,and the inhibitory rates were 18.6% and 32.8.The ginsenoside Rh1 powder was dissolved with dimethyl sulfoxide(DMSO).The following experiments were divided into three groups: blank control group,ginsenoside Rh1 group.he results of flow cytometry showed that ginsenoside Rh1 could induce apoptosis of SKBR3 cells and affect the proliferation of SKBR3 cell cycle at S phase.2.Effects of ginsenoside Rh1 on migration and invasion of HER2 positive breast cancer cells.The results of scratch test showed that ginsenoside Rh1 inhibited the migration of SKBR3 cells after 48 h.The results of Transwell invasion test showed that the invasive ability of SKBR3 cells was significantly decreased after 24 h intervention by ginsenoside Rh1.3.The results of real-time quantitative PCR RT-PCR showed that ginsenoside Rh1 had no effect on the expression of DDX5 gene in SKBR3 cells.Western-blot analysis showed that the expression of DDX5 protein in human breast cancer SKBR3 cells could be reduced by 500 u Mol ginsenoside Rh1.4.By collecting the detailed pathological data and clinical staging of breast cancer tissue sections for DDX5 immunohistochemical staining experiment and score,statistical analysis showed that HER2 positive breast cancer tissue DDX5 expression is higher,and the high expression of DDX5 in patients with poor pathological grade.The clinical stage was late.Conclusion:In this study,we found that ginsenoside Rh1 could inhibit the proliferation of HER2 positive breast cancer cells(SKBR3),induce apoptosis of SKBR3 cells,block them in S1 phase,inhibit the migration and invasion of SKBR3 cells.The expression of DDX5 protein in SKBR3 cells was inhibited by immunohistochemical staining.The results of immunohistochemical staining indicated that the expression of DDX5 in HER2 positive breast cancer tissues was higher than that in patients with high expression of DDX5,and the expression of DDX5 was lower than that of patients with high expression of DDX5 in clinical stage and pathological grade of breast cancer.Clinical staging later.DDX5 may become HER2 positive Ginsenoside Rh1 may inhibit the expression of DDX5 protein by inhibiting the post-transcriptional process of DDX5 and thus affect the malignant biological behavior of breast cancer SKBR3 cells.