| Background:Tongue Squamous Cell Carcinoma(TSCC)is a malignant tumor of the oral cavity with a high degree of malignancy and is easy to early metastasis.Smoking and drinking are the main risk factors of tongue squamous cell carcinoma.Surgical treatment is the main treatment,sometimes adjuvant therapy such as radiotherapy and chemotherapy is needed to improve the survival rate of patients.However,surgical treatment will cause maxillofacial tissue defects and reduce the quality of life of patients.Therefore,it is very necessary to study some drugs or gene therapy with less damage to patients.Clinically,only a few drugs can be used to treat tongue cancer.In recent years,it has been found that ginsenoside Rd has anticancer effect.This study mainly analyzed the inhibitory effect of ginsenoside Rd on tongue cancer and its mechanism.Objective:The main purpose of this study is to explore the inhibitory effect of ginsenoside Rd on tongue cancer and its mechanism.So as to provide basis for ginsenoside Rd as a potential drug in the treatment of tongue cancer.Methods:1.CCK8 experiment was used to study the effect of different concentrations of ginsenoside Rd on the proliferation of TSCC.Flow cytometry was used to study the apoptosis.The effect of ginsenoside Rd on the migration ability of TSCC cells was detected by the wound-healing assay.Transwell was used to detect the effect of ginsenoside Rd on the invasion of TSCC cells.Colony formation assays was used to detect the effect of ginsenoside Rd on colony formation ability of TSCC.2.H19 and miR-675-5p expressions in tongue cancer tissue were analyzed by qPCR.After overexpression or knockdown of H19 and miR-675-5p,Transwell assay was used to assess invasion;wound-healing assay to assess migration;colony formation assays to test the ability of cells to form colonies;and CCK8 to study growth.3.CDH1 expression in tongue cancer tissue was analyzed by qPCR.E-cadherin expression was detected using western blot.CRISPR/Cas9 system was used for CDH1 knockdown.After overexpression or knockdown of CDH1,Transwell assay was used to assess invasion;wound-healing assay to assess migration;colony formation assays to test the ability of cells to form colonies;and CCK8 to study growth.4.H19,miR-675-5p and CDH1 expressions were analyzed by qPCR after ginsenoside Rd treatment.And E-cadherin expression was detected using western blot after ginsenoside Rd treatment.Bioinformatics analysis was used to study the relationship between miR-675-5p and CDH1.When we overexpress or knockdown H19,miR-675-5p and CDH1 expressions were detected by qPCR,and E-cadherin expression was detected using western blot.When we overexpress or knockdown miR-675-5p,CDH1 expression was analyzed by qPCR,and E-cadherin expression was detected using western blot.Results:1.CCK8 assay showed that ginsenoside Rd could inhibit the growth of SCC9 cells.Meanwhile,flow cytometry showed that ginsenoside Rd could increase the apoptosis of SCC9 cells.The wound healing assay and Transwell assay showed that ginsenoside Rd could inhibit the migration and invasion of SCC9 cells.2.H19 was highly expressed in tongue cancer.The wound healing assay and Transwell assay showed that H19 could promote the migration and invasion of SCC9 cells.Colony formation assay showed that H19 could increase the colonies of SCC9 cells.CCK8 experiment showed that overexpression of H19 had no significant effect on the proliferation of SCC9 cells,while knockdown of H19 could inhibit the proliferation of SCC9 cells.3.qPCR analyses showed that miR-675-5p expression was higher in tongue cancer tissues than in normal tissues.The wound healing assay and Transwell assay showed that miR-675-5p could promote the migration and invasion of SCC9 cells.In the colony formation assay,miR-675-5p could increase the colonies of SCC9 cells.CCK8 assay showed that miR-675-5p could enhance the proliferation of SCC9 cells.4.In contrast to H19 and miR-675-5p,the expression of CDH1 was low in tongue cancer.The wound healing assay and Transwell assay showed that CDH1 could inhibit the migration and invasion of SCC9 cells.Colony formation assay showed that CDH1 could inhibit the colony formation ability of SCC9 cells.CCK8 assay showed that CDH1 could inhibit the proliferation of SCC9 cells.5.After SCC9 cells were treated with ginsenoside Rd,qPCR results showed that the expression of H19 and miR-675-5p reduced,while the expression of CDH1 increased.Western Blot showed that the expression of E-cadherin increased.H19 can increase the expression of miR-675-5p and inhibit the expression of CDH1 and Ecadherin.Mi R-675-5p can inhibit the expression of CDH1 and E-cadherin.Conclusion:Ginsenoside Rd can inhibit the expression of H19 and miR-675-5p and increase the expression of CDH1.When the expression of CDH1 increases,the expression of E-cadherin increases,which makes the connection between cells tighter,thus inhibiting the migration and invasion of tongue cancer cells.Therefore,ginsenoside Rd inhibits the migration and invasion of tongue cancer cells by regulating the H19/miR-675-5p/CDH1 axis. |
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