Expressions Of NuSAP1in Hepatocellular Carcinoma And Its Correlations With Hepatocellular Carcinoma Recurrence

[Backgrounds]Hepatocellular carcinoma (HCC) is one of the most common and frequent malignancies. Its high rate of recurrence after surgical resection is one of the key reasons for the low survival rates and a poor prognosis. However, the cause and the machenism of HCC remains unexplored. To improve patient prognosis, it is very important to elucidate the biological mechanisms that control the tumor-initiating and development and predict the early recurrence of HCC. Nucleolar-spindle associated protein1(NuSAP1) is a microtubule-associated protein that plays a central role in spindle assembly and act as a key regulator to warrant the process of cell division.The levels of NuSAP1expression were strictly regulated and dynamically changed during different stages of the cell cycle. The transcription factor E2F family play a key roles in cotrolling the cell cycle. The binging of E2F1to its binding site which located on the promoter of NuSAP1was able to markedly promote the NuSAP1transcripton. Recent studies indentify overexpression of NuSAP1in many other tumors,and the expression level of NuSAP1seem to be related to the invasiveness and prognosis of the tumors.But whether the same mechanism exsit in HCC has not been report. [OBJECTIVE]Owing to the important function of the NuSAP1in the cell cycle,the disturb of its function may associated with the development and recurrence of cancer. The aims of the research are mainly to investigate the expression level of NuSAP1in HCC cell lines and HCC tissues,elucidate its clinical significances and its relationship with E2F1,explore its role in occurrence, development and recurrence of hepatocellular carcinoma.[METHODS](I).The expression level of NuSAP1and its mRNA, E2F1protein in tumor tissues and non-tumor tissues from patients with HCC was detected by semi-qRT-PCR, quantitative real-time PCR and immunohistochemical; the relationship between its expression and the clinicopathologic features was studied respectively. Then the correlations between NuSAP1and E2F1in HCC were studied.(Ⅱ). HCCLM3cell line with high aggressive phenotype was choose, After inhibited with siRNAs, NuSAP1mRNA and protein expression level were detected by quantitative real-time PCR and Western blotting.Cell proliferation activity was analysised by the method of MTS and Edu,Cell migration and aggressive capability was tested by the method of transwell.Flow cytometry was used to determined cell cycle and apoptosis, respectively.(Ⅲ). After inhibited the expression of E2F1in HMHCC97H cell line with siRNAs, NuSAP1mRNA and protein expression level were detected by quantitative real-time PCR and Western blotting(analysis among the inhibite-before and inhibite-after HCC cell lines),and then their relationship between E2F1and NuSAP1in hepatocellular carcinoma were studied.[RESULTS]1.There were apparently higher expressions of NuSAP1and its mRNA in HCC tumor tissues than in non-tumor tissues(P<0.05).With univariate analysis showed that the expression of NuSAP1and its mRNA in HCC was associated with TNM classification, BCLC classification, lymphatic metastasis, early recurrence, tumor thrombi and histological differentiation (P<0.05). Meal while, the expression of NuSAP1mRNA was also associated with the serum AFP level;The expression of NuSAP1was also associated with tumor size and liver cirrhosis.With multivariate analvsis, NuSAPl is a independent risk factor for early recurrence (P=0.005).Survival analysis showed that the expression of NuSAP1in HCC was associated with postoperative recurrence (X2=5.939, P=0.015). The surival time without tumor was positively correlated with NuSAP1protein expression (Kaplan-Meier, log-rank test, P<0.05).2.The expressions of E2F1protein were apparently higher in tumor tissues than in non-tumor tissues(P<0.05). The expression of E2F1protein in HCC was associated with histological differentiation, early recurrence, BCLC classification and TNM classification (P<0.05), but not to sex, age, tumor thrombi, tumor size, liver cirrhosis, lymphatic metastasis,tumor number, AFP level,HBV and the presence of tumor encapsulation (P>0.05). Survival analysis showed that the expression of E2F1protein in HCC was associated with postoperative recurrence (X2=8.952, P=0.003). The surival time without tumor was positively correlated with E2F1protein expression (Kaplan-Meier, log-rank test, P<0.05). There was positive correlation between the expression of E2F1protein and NuSAPl in tumor tissues (P<0.05);3.1n the study of inhibited HCC cell lines,inhibited the expression of NuSAP1could apparently decrease the cell numbers,cell proliferation activity,cell migration capability and cell aggressive capability,but increase the cell cycle G2/M transform and cell apoptosis(P<0.05); To inhibite the expression of E2F1protein could apparently decrease the expression of NuSAP1(P<0.05).[CONCLUSION]1、The expression of NuSAP1in HCC tissues was higher than that in non-tumor tissues, and correlated with postoperative early recurrence, Higher expression of NuSAP1might lead to a poor prognosis.2, The expression of E2F1protein in HCC tissues was higher than that in non-tumor tissues, and positive correlated with the level of NuSAPl,this indicate E2F1protein maybe act as a regulator to NuSAP1.3、NuSAPl maybe a oncogene in HCC cell lines.,its high expression maybe take part in HCC angiogenesis and development.4、E2F1maybe act as a regulator to NuSAPl in HCC cell lines.