| BackgroundPrimary liver cancer is a popular malignant tumor of digestive system worldwide,which has hidden onset,rapid progress,easy metastasis and poor prognosis.Hepatocellular carcinoma(HCC)is the most predominant pathological type in clinic,and its occurrence and progression are closely related to hepatitis B virus(HBV).At present,the gold standard for the diagnosis of HCC is still the results of histopathology and cytology.In view of its trauma and high risk,the clinical diagnosis of HCC is usually done by combining imaging examination with serum AFP level.In consideration of the limitation of the existing methods,it is urgent to search a new non-invasive biomarker to break the current shackles.Bone morphogenetic protein 2(BMP2)and bone morphogenetic protein 10(BMP 10)belong to the bone morphogenetic proteins family and can effectively regulate cell differentiation,proliferation and apoptosis,further participating in the regulation of embryonic development and the dynamic balance of various tissues and organs after birth.Studies have shown that BMPs may regulate the growth,differentiation or apoptosis of many types of cancers.They can inhibit tumor growth and metastasis,but on the contrary,they can also be used as tumor promoters by activating oncogenes,stimulating tumor microenvironment and other mechanisms.BMP2 has been proved to be expressed in tumor tissues and is related to the occurrence,development,metastasis and prognosis of several kinds of malignant tumors.However,the contradictory results suggest that the role of BMP2 in HCC is not clear,which may be in connection with the types of HCC cell lines and the microenvironment of tumor cells.Presently,the expression of BMP2 in peripheral blood mononuclear cells(PBMCs)of patients with HCC and its relationship with HCC have not been reported.A large number of experiments in vitro have shown that BMP 10 can inhibit the growth of a variety of cancer cells and is relevant to the invasion and migration of cancer cells,suggesting that BMP10 may act as a tumor suppressor in different types of cancer.Recent studies have found that the expression of BMP 10 is down-regulated in HCC,indicating that BMP 10 has a potential tumor inhibitory effect in HCC.Whereas,the expression level of BMP10 in PBMCs of patients with HCC and its impact on tumor growth and metastasis need to be further explored.The study is designed to detect the expression levels of BMP2 and BMP 10 mRNA in PBMCs of HCC patients,chronic hepatitis B(CHB)patients and healthy controls(HCs),so as to explore their potential diagnostic value in HBV-related HCC.Methods220 samples were included in the study totally,covering 105 patients with HCC,85 patients with CHB and 30 HCs.The expression levels of BMP2 and BMP 10 mRNA in three groups of PBMCs were detected by real-time quantitative polymerase chain reaction(RT-qPCR).Results1.The expression level of BMP2 mRNA in HCC patients was apparently greater than that in CHB group(P<0.001)and HCs(P<0.001).Furthermore,The BMP2 mRNA level in CHB patients also exceeded the level in HCs(P<0.007).Besides,BMP 10 mRNA concentrations in HCC group was obviously lower than that in CHB group(P<0.001)and HCs group(P<0.001),whereas there was no difference in BMP 10 mRNA expression level between CHB group and HCs group(P<0.852).2.The mRNA expression of BMP2 in HCC group was positively correlated with TBA(r=0.221,P=0.024),international normalized ratio(INR)(r=0.222,P=0.023),and HBsAg(r=0.241,P=0.013),but has nothing to do with age,PLT,ALT,AST,TBIL,ALB,PTA,Cr,HBV DNA,HBeAg,Child-Pugh grade,tumor size,vascular invasion,lymphatic metastasis and TNM stage.In addition,the BMP2 mRNA level in HCC patients with multiple tumors and with ascites was significantly higher than that in patients with single tumor and without ascites.(Z=2.962,P=0.003;P=0.033,Z=-2.137).3.In HCC patients,the level of BMP 10 mRNA was negatively correlated with TBA,INR,Cr and PTA,but not with age,PLT,ALT,AST,TBIL,albumin,HBeAg,ascites,Child-Pugh grade,tumor size and number of tumors.The expression level of BMP10 mRNA was fallen in vascular invasion(+)and lymphatic metastasis(+)group compared with vascular invasion(-)and lymphatic metastasis(-)group(P=0.004,Z=-0.284;P=0.006,Z=-2.752).Moreover,BMP 10 mRNA average level of the advanced HCC was lower than that in the early HCC(p=0.002,Z=-3.105).4.When distinguishing HCC and CHB,the AUC of BMP2 and BMP10 mRNA was 0.751 and 0.794 respectively,which was not apparently different from AUC 0.715 of AFP(P=0.473;P=0.090).The cut-off of BMP2 in the diagnosis of HCC was 0.0224,of which sensitivity and specificity were 84.8%and 56.5%respectively.The cut-off of BMP10 was 0.127,and its sensitivity and specificity were 71.4%and 77.6%respectively.However,when any of the three combined to diagnose HCC(BMP2 with BMP10,BMP2 with AFP,BMP10 with AFP),AUC were 0.844,0.808,0.831 respectively.There were significant differences compared with AFP alone in the diagnosis of HCC(P=0.004,0.012,0.0035,respectively).In addition,the AUC of the combination of BMP2,BMP10 and AFP was 0.873,which was obviously rising compared to that of any combination of the two and the single index(P<0.001).5.For patients with negative AFP(AFP<20 ng/ml),the AUC of BMP2 and BMP10 mRNA in the diagnosis of HCC was 0.764 and 0.814,and the AUC of combinating BMP2 with BMP 10 to diagnose HCC was 0.863.The sensitivity and specificity of BMP2 mRNA cut-off were 81.1%and 60.3%respectively,that of BMP10 mRNA were 70.3%and 81.0%and of combination were 73.0%and 93.7%.6.Multivariate logistics regression analysis indicated that age>50 years old,AFP>20 ng/ml,BMP2 mRNA>0.0224,BMP 10 mRNA<0.127 and INR level may be the independent risk factors for HBV-related HCC.Conclusions1.The expression level of BMP2 mRNA in PMBCs of HCC patients was significantly higher than that of CHB patients and HCs,while the BMP 10 mRNA level of HCC group was greatly lower than that of CHB group and HCs.2.The expression level of BMP2 mRNA in patients with multiple tumors was markedly increasing than that in patients with single tumor,indicating that BMP2 mRNA may be related to the intrahepatic metastasis of HCC.The BMP2 mRNA level in patients with HCC was positively correlated with TBA,INR and ascites,suggesting that the up-regulation of BMP2 expression may be linked to the severity of the disease.3.The expression level of BMP 10 mRNA in advanced HCC subgroup,patients with vascular invasion and lymph node metastasis was dramatically decreasing compared to that in early HCC subgroup,patients without vascular invasion and lymph node metastasis,showing that BMP 10 may be related to tumor progression and metastasis.In HCC group,BMP10 mRNA was negatively correlated with TBA,INR,Cr and positively correlated with PTA.It can be speculated that the down-regulated expression of BMP10 in patients with HCC may reflect the severity of HCC.4.In the diagnosis of HCC,the value of BMP2 and BMP 10 combined with AFP is significantly higher than that of the three alone,and the diagnostic accuracy of the combination of the three is the highest.BMP2 and BMP 10 still have pretty diagnostic significance for HCC patients of negative AFP.This implies that BMP2 and BMP 10 may be used as non-invasive serum biomarkers for the diagnosis of HCC,which puts forward a new approach for the diagnosis of HCC. |
PDF Full Text Request