| BackgroundLiver cancer is one of the most common malignant tumors in the world.The incidence and mortality of liver cancer in China are still on the rise.The annual incidence and mortality of liver cancer in China are very similar.This shows that the existing diagnosis and treatment strategies and measures to reduce the total mortality of liver cancer in five years are very limited.This situation shows that there is an urgent need to move forward the prevention and treatment strategy of liver cancer.Chinese medicine in the treatment of precancerous lesions of liver cancer has obvious advantages in controlling the disease and preventing cancerization.To carry out the study of precancerous lesions of liver cancer with Chinese medicine is of great significance to reduce the incidence of liver cancer.ObjectiveThis study was aimed to clarify the clinical efficacy of Kangxian-Yiai formula(KXYA)in the treatment of precancerous lesions of liver cancer,and to explore the significance of the expression of mammalian target of rapamycin(mTOR),hypoxia inducible factor1α(HIF-1α)and vascular endothelial growth factor(VEGF)in the process of malignant transformation of hepatocytes(from liver cirrhosis to precancerous lesions,and then to hepatocellular carcinoma).Moreover,based on the hypoxia microenvironment and angiogenesis-related genes,we aimed to reveal the regulatory effect of KXYA on precancerous lesions through mTOR/HIF-1α/VEGF signal pathway.Through this study,in order to provide a strong basis for further clinical application of the KXYA.Methods1.Observation on the efficacy of KXYA in the treatment of precancerous lesions of liver cancer.The data of 44 patients with precancerous lesions in Dongzhimen Hospital affiliated to Beijing University of Chinese Medicine from January 2008 to January 2019 were collected retrospectively.All the patients were treated with KXYA for more than half a year.The basic information of patients and baseline laboratory examination were investigated and analyzed,and the changes of precancerous nodules(including canceration rate and nodule stability rate)in patients with precancerous lesions were observed during the treatment of KXYA,and the effects of baseline indexes on clinical outcome of patients with precancerous lesions were analyzed.2.The expression and significance of mTOR,HIF-1α and VEGF in precancerous lesions and hepatocellular carcinoma were analyzed based on data mining.The expression data of MTOR,HIF1 A and VEGFA genes in hepatocellular carcinoma,precancerous lesions,cirrhotic tissues and normal tissues were extracted from Oncomine and TCGA database,and the expression of each gene in different disease stages was analyzed.Kaplan-Meier Plotter online database was used to analyze the relationship between prognosis and the expression of MTOR,HIF1A and VEGFA.To explore the significance of liver tissue transcription of mTOR,HIF-1αand VEGF i n the process of malignant transformation of hepatocytes3.To explore the expression and significance of mTOR,HIF-lα and VEGF in the process of malignant transformation of hepatocytes based on clinical cases.Three groups of patients with liver cirrhosis,precancerous lesions and primary liver cancer were included.The serum of the patients was collected and separated.The contents of mTOR,HIF-1 a and VEGF in the serum of the three groups were detected by enzyme-linked immunosorbent assay(Elisa).At the same time,the alpha fetoprotein(AFP)and biochemical indexes of the three groups were collected,and the indexes of the three groups were compared and analyzed.The purpose of this study is to explore the significance of serum levels of mTOR,HIF-1α and VEGF in the process of malignant transformation of hepatocytes.4.The regulatory effect of KXYA on precancerous lesions of liver cancer through mTOR/HIF-lα/VEGF signal pathway.The rat model of precancerous lesion of liver cancer was established by diethylnitrosamine.The rats were intervened at the 9th week after the establishment of the model for 6 weeks,and the liver tissue was taken at the end of the 14th week.The pathological changes of liver tissue were observed by HE and Masson staining.The expression of glutathione-S-transferases-π(GST-Pi)in precancerous lesions was detected by immunohistochemistry and Western blot to determine the degree of precancerous lesions.Western blot and real-time fluorescence quantitative PCR were used to detect the expression of glucose transporter-1(GLUT1),pyruvate kinase M2(PKM2),mTOR.HIF-1α and VEGF protein and mRNA level.Results1.Observation on the efficacy of KXYA in the treatment of precancerous lesions of liver cancer.A total of 44 patients with precancerous lesions were included in this study,including 38 males(86.4%)and 6 females(13.6%).The average age of female patients was 66 years old(range:58 to 73 years),and the male patients was 50 years old(range:29 to 77 years).The age of female patients was significantly higher than that of males(P=0.003).In terms of outcome,precancerous nodules remained stable(14 cases)or smaller(18 cases)in 32 patients(72.7%),and enlarged(9 cases)or cancerous(3 cases)in 12 cases(27.3%).In terms of the influence of clinical baseline index on clinical outcome,the neutrophil count in precancerous nodule enlarged or cancerous group was significantly lower than that in small or stable group(P=0.047),and the level of aspartate aminotransferase(AST)in precancerous nodule enlarged or cancerous group was significantly higher than that in small or stable group(P＝0.032).2.To explore the expression and significance of mTOR,HIF-lα and VEGF in the process of malignant transformation of hepatocytes based on clinical cases.A total of 67 patients were included in this study,including 20 patients with liver cirrhosis,23 patients with precancerous lesions and 24 patients with hepatocellular carcinoma.The age of patients in liver cancer group was older than that in liver cirrhosis and precancerous lesions group(P<0.01).During the malignant transformation of hepatocytes,the levels of AFP,glutamic oxaloacetic transaminase.y-glutamyl transpeptidase,direct bilirubin and serum creatinine increased gradually and reached the peak in the stage of hepatocellular carcinogenesis,while the levels of albumin,cholinesterase,prothrombin time and prothrombin activity decreased gradually and reached the lowest in the stage of hepatocellular carcinoma.The serum levels of mTOR.HIF-lα and VEGF increased gradually during the malignant transformation of hepatocytes in the three groups.The levels of serum mTOR,HIF-1α and VEGF in liver cancer group were significantly higher than those in liver cirrhosis group(P<0.01).3.The expression and significance of mTOR,HIF-1α and VEGF in precancerous lesions and hepatocellular carcinoma were analyzed based on data mining.Through the analysis of Oncomine and TCGA database,it was found that most cohorts showed that MTOR,HIF1A and VEGFA were highly expressed in liver cancer tissues in terms of gene differential expression between liver cancer and normal tissues.In terms of gene differential expression between precancerous lesions and liver cancer tissues,MTOR was highly expressed in liver cancer tissues.while VEGFA gene was highly expressed in precancerous lesions.There was no significant difference in the expression of HIFlA between precancerous lesions and liver cancer tissues.In terms of prognosis,the prognosis of patients with high expression of HIFIA and VEGFA genes was poor(P<0.05).There was no significant difference in prognosis between patients with high expression of MTOR and low expression group(P>0.05).4.The regulatory effect of KXYA on precancerous lesions of liver cancer through mTOR/HIF-1α/VEGF signal pathway.KXYA can significantly improve the pathological changes of precancerous lesions induced by diethylnitrosamine in rats,and reduce the protein expression of PKM2 and GLUT-1,and significantly down-regulate the mRNA and protein expression of mTOR,HIF-1α and VEGF in liver tissueConclusions1.KXYA can effectively reduce the incidence of liver cancer in the treatment of precancerous lesions of liver cancer.2.Through the verification of data mining and clinical cases,it was found that mTOR,HIF-1α and VEGF were closely related to the malignant transformation of hepatocytes3.KXYA can inhibit the expression of glycolysis-related genes PKM2 and GLUT-1,and can inhibit the mTOR/HIF-lα/VEGF pathway,which may be one of its mechanisms for the treatment of precancerous lesions of liver cancer. |
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