| Background:Hepatocellular carcinoma (HCC) is one of the modalities of malignant tumors.It had belong to the medium and late period when HCC was diagnosised.It also bringed certain difficulty radical cure for HCC because of the invasion and metastasis of tumor cells, apoptosis escape, angiogenesis and tumor drug resistance complex biological behavior. Therefore,it has important theoretical and clinical application value further study if these complex biological malignant phenotype of signal transmission and the adjustment mechanism can be researched. It can provide new targets for the diagnosis and treatment of HCC. Jidesheng Sheyao has many effects such as clearing heat, relieve swelling and pain, promoting circulation and removing stasis. It was mainly used to treat snake wound. In recent years, It was also used to cure liver cancer in clinical practice and got certain curative effect.Therefore, this study can provide new ideas for HCC diagnosis, pathogenesis and treatment, in addition,it can also provide new intervention targets for HCC treatment.Objective:Research the expression of miR-335 in hepatocellular carcinoma and the effects of Jidesheng Sheyao medicated serum on human hepatocellular carcinoma cell HuH-7.Methods:The real-time quantitative polymerase chain reaction (PCR) was used to detective the expression level of miR-335 in the serum of HCC patients and healthy controls. The relationship of their expressions with HCC patients as well as clinical pathological parameters were further analyzed. The real-time quantitative polymerase chain reaction (PCR) was used to detective the expression level of miR-335 in cancer patients and hepatoma cell lines. The effect of miR-335 on HuH-7 cells proliferation was tested by CCK-8 assay. The effect of miR-335 on potential target protein Bcl-w was tested by Western-blot. The effect of Jidesheng Sheyao medicated serum on HuH-7 cells proliferation was tested by CCK-8 assay. qRT-PCR was used to detective the regulation function of miR-335 on human hepatocellular carcinoma cell HuH-7.Results:Result showed that the relative miR-335 expression was significantly lower in the serum of HCC than that in the N C groups (P<0.01). The relative miR-335 expression in the serum of HCC with gender, age, the tumor size, TNM stage, lymph node metastasis and AFP were not correlated (P>0.05). By comparison, it with cancer embolus were positively correlated (P<0.01). ROC analysis showed that the critical value of miR-335 was 1.28, with sensitivity of 78%, specificity of 70%, and area under the curve of 0.75. The best critical value of AFP, with sensitivity of 76%, specificity of 97%, and AUC of 0.83. The combination of serum miR-335 and AFP, with sensitivity of 84%, specificity of 97%, and AUC of 0.93. The relative miR-335 expression had obvious difference between cancer and cancer adjacent tissues (P<0.05). The relative miR-335 expression in the liver cancer with gender, age, the tumor size, TNM stage, lymph node metastasis and AFP were not correlated (P>0.05). By comparison, it with cancer embolus were positively correlated (P<0.05). Cell screening results showed that miR-335 has significantly lower expression in liver cancer HuH-7 cells. Up-regulation of miR-335 may suppress the cellular proliferation and induce cell apoptosis in HuH-7 cells, at least in part, through down-regulation the expression level of target gene Bcl-w. Jidesheng Sheyao medicated serum may suppress the cellular proliferation and regulate miR-335 on human hepatocellular carcinoma cell HuH-7.Conclusions:Lower expression of serum miR-335 in HCC patients is expected to become a new kind of biomarkers for the diagnosis of liver cancer. The combination of serum miR-335 and AFP can elevate the diagnostic efficiency for HCC.The relative miR-335 expression has obvious difference between cancer and cancer adjacent tissues. By comparison, it with cancer embolus was positively correlated. Up-regulation of miR-335 may suppress the cellular proliferation and induce cell apoptosis in HuH-7 cells, at least in part, through down-regulation the expression level of target gene Bcl-w. At last, miR-335 may serve as a novel molecular target for treatment of liver cancer. Jidesheng Sheyao is expected to become new drug treatment for liver cancer. |
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