| Background and object:Esophageal squamous cell carcinoma(ESCC)is one of the most common human malignancies and has a high mortality rate.Men are more common than women.The morbidity and mortality of esophageal squamous cell carcinoma(ESCC)all over the world have been increasing year by year.Although the diagnosis and treatment of esophageal cancer have been improved in recent ten years,the prognosis of esophageal squamous cell carcinoma is still poor.At present,many risk factors for esophageal cancer have been identified,but the mechanisms by which they cause disease are not yet known.Micro RNAs(mi RNAs or mi Rs),small ribonucleic acids,are small endogenous,non-coding RNAs approximately 18 to 24 nucleotides in length that bind to the 3’untranslated region of the target gene m RNA by base-pairing pairing(3 ’-UTRs),resulting in the inhibition of target messenger RNA or protein synthesis,thus regulating gene expression at post-transcriptional level.A large number of studies have confirmed that the development of cancer is related to the abnormal expression of mi RNAs.As tumor suppressor genes and / or proto-oncogenes,mi RNAs are involved in the regulation of proliferation,apoptosis,metastasis and invasion of tumor cells.Recent studies have shown that mi R-218 affects tumorigenesis and progression by inhibiting the proliferation and invasion of tumor cells,such as mi R-218,which regulates the proliferation and metastasis of an esophageal squamous cell carcinoma of the ROBO1 and BMI1 genes through targeting.In addition,mi R-218 promotes esophageal squamous cell carcinoma chemosensitivity to cisplatin.However,whether mi R-218 inhibits the proliferation and metastasis of esophageal squamous cell carcinoma cannot be clarified by acting on a new target gene.This study aimed to discuss the biological functions of mi R-218 in inhibiting the proliferation,metastasis and invasion of esophageal squamous cell carcinoma by targeting Hif-1α.Research methods:1.80 cases of surgical resected esophageal squamous cell carcinoma tissue and adjacent normal tissue specimens were collected,and real-time fluorescence quantitative PCR method was used to detect the expression of esophageal squamous cell carcinoma cell line and normal esophageal mucosa epithelial cells,cancer tissue specimens and adjacent normal tissue specimens the expression of mi R-218 was detected,and the differential expression of mi R-218 was statistically analyzed.The correlation between the differential expression of mi R-218 and the clinicopathological characteristics and survival prognosis was also analyzed.2.The effect of mi R-218 overexpression on the proliferation and invasiveness of esophageal squamous cell carcinoma cells was examined to determine the biological function of mi R-218.3.The bioinformatics website was used to predict the potential target gene of mi R-218,HIF-1α,and to verify whether mi R-218 can regulate Hif-1α(HIF-1α)in vitro and affect the proliferation and invasion of esophageal squamous cell carcinoma cells.Research result:1.To detect the expression of mi R-218 and HIF-1α in esophageal squamous cell carcinoma,adjacent normal tissue and various squamous cell carcinoma cell lines and normal esophageal epithelial cells:1.1 The expression of mi R-218 in esophageal squamous cell carcinoma and adjacent normal tissues was detected by real-time fluorescence quantitative PCR: mi R-218 was relatively low expression in esophageal squamous cell carcinoma tissues compared with adjacent normal tissues;the relative expression level of mi R-(T = 4.364,p = 0.032),lymph node metastasis(t = 17.875,p <0.01)and TNM stage(t = 3.279,p = 0.002)(T =15.201,p <0.01);1.2 The expression of mi R-218 and HIF-1α in esophageal squamous cell carcinoma cell lines was detected by real-time fluorescence quantitative PCR.Compared with normal esophageal epithelial cells(HET-1a),esophageal squamous cell carcinoma cell lines KYSE30,KYSE70,KYSE150,In KYSE450 and KYSE510,mi R-218 was relatively low expression;in contrast,HIF-1α was relatively high expression.2.Mi R-218 overexpression in esophageal squamous cell carcinoma cell line KYSE30 proliferation,migration and invasion: compared with the NC control group,mi R-218 mimics group 0h,24 h,48h,72 h cell proliferation was significantly inhibited;mi R-218 mimics group increased the rate of cell apoptosis;mi R-218 mimics group cells decreased migration and invasion.The difference was statistically significant,P <0.05.3.Luciferase reporter assay was used to verify the role of mi R-218 in the regulation of HIF-1α expression.4.HIF-1α gene expression was silenced by si RNA to observe the effects of proliferation,migration and invasion of KYSE30 cells:4.1 The si RNA was transfected into KYSE30 cells by transient transfection and the interference efficiency was verified by real-time fluorescence quantitative PCR and Western blot.The down-regulation of Hif-1α gene expression in si RNA-3 group was the most significant,P < <0.05;4.2 The results showed that compared with the NC group,the proliferation,metastasis and invasion ability of si RNA-3 group were significantly decreased,the difference was statistically significant,P <0.05conclusion:1.Compared with the normal tissue samples adjacent to esophageal cancer,mi R-218 is low expressed in esophageal squamous cell carcinoma tissues;similarly,mi R-218 is low expressed in many esophageal squamous cell lines with esophageal normal mucosa epithelial cells.2.Overexpression of mi R-218 can inhibit the proliferation and invasion of esophageal squamous cell carcinoma cells.3.Hypoxia-inducible factor-1α(HIF-1α)is the target gene of mi R-218.Overexpression of mi R-218 inhibits the expression of HIF-1α protein.Silencing the expression of HIF-1α gene can inhibit the proliferation of esophageal squamous cell carcinoma cells Invasive ability,and this effect is similar to overexpression of mi R-218. |
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