1?the Role And Molecular Machanism Of Nuclear Receptor Coactivators 6(NCoA6)in Tumorigenesis And Progression Of Hepatocellular Carcinoma 2?laparoscopic Right Hepatectomy For Hepatocellular Carcinoma:a Propensity Score Matching Analysis Of Outcomes Compar

Primary hepatocellular carcinoma(HCC)is one of the most frequent cancers worldwide,especially in China where the morbidity and mortality accounted for more than half of the total number around the world.HCC is a complex disease with abnormal changes in molecular mechanisms caused by multiple risk factors.Although comprehensive treatment based on hepatectomy has made some progress,the overall prognosis of patients is still poor.Therefore,finding ways for early diagnosis and treatment of HCC is an important method to improve the prognosis.Nuclear receptor coactivators 6(NCoA6)is a multifunctional co-regulator or co-activator necessary for transcriptional activation of a wide spectrum of target genes including nuclear hormone receptors(NRs)and several important transcription factor.NCoA6 is a pleiotropic co-regulator involved in growth,development,wound healing and maintenance of energy homeostasis,etc.Research reported that the NCoA6 gene is amplified and overexpressed in numerous malignancies like breast cancer,colorectal cancer,lung cancer,gastric cancer,pancreatic ductal adenocarcinoma,and melanoma etc.,whereas the expression and function of NCoA6 in HCC has not been fully investigated.In the part one of this study,we tested the expression of NCoA6 in surgical samples from HCC patients and HCC cell lines,combining with a series of experiments in vivo and in vitro to find the potential role and molecular mechanism of NCoA6 in tumorigenesis and progression of HCCAs mentioned above,surgery is still the most important component of comprehensive treatment for HCC.However,severe surgical trauma is inevitable in the conventional open surgery.Minimally invasive surgery,typically characterized by laparoscopic approach,is one of the main surgical advances in the twenty-first Century A growing number of publications have described the feasibility,safety,and adequacy of laparoscopic hepatic procedures.However,majority of HCC patients have underlying liver diseases,such as liver cirrhosis and viral hepatitis,which inevitably damage liver function making them easy to bleed and poor tolerance to surgery.If the tumor is located in the right liver,when hemihepatectomy is necessary,finishing this challenging procedure under laparoscopy is difficult and risky.Therefore,few studies have been reported using laparoscopic right hepatectomy(LRH)for HCC patients.In the part two of this study,we summarized the operative experience of LRH performed in our center and evaluated the feasibility and safety of this procedure by comparing its short-and long-term clinical outcomes with those of open right hepatectomy(ORH)through a propensity score-matched(PSM)analysis to minimize selection biasPart one:the role and molecular mechanism of nuclear receptor coactivators 6(NCoA6)in tumorigenesis and progression of hepatocellular carcinomaSection one:the expression and its clinical significance of NCoA6 in hepatocellular carcinomaMethods Databases like Oncomine and TGCA etc.were searched,and the expression of NCoA6 in HCC and its correlation with prognosis were analyzed by bioinformatics.NCoA6 expression in HCC cell lines and human HCC tissues was evaluated by PCR,Western blot,and immunohistochemical staining to analyze the correlation between NCoA6 expression and clinicopathological features and prognosis of patients.Results Biological analysis showed that NCoA6 was significantly elevated in HCC and had a significant correlation with prognosis.The results of PCR and Western blot confirmed that NCoA6 was highly expressed in HCC cell lines and clinical HCC tissues.The immunohistochemical analysis of a cohort containing 103 HCC patients,combining with clinical and pathological features of HCC patients demonstrated that the high expression of NCoA6 is closely related to the tumor size(p=0.004).Kaplan-Meier analysis showed that a high level of NCoA6 expression was correlated with poor overall and disease-free survival of HCC patients(p=0.01).The results of univariate and multivariate analysis indicated that NCoA6 over-expression was significantly correlated with overall and disease-free survival of HCC patients(p<0.05).Conclusions The NCoA6 expression was significantly upregulated in tumor cell lines and tissues relative to normal hepatic tissues,which was also significant correlation with the prognosis of patients.Section two:the function of NCoA6 in hepatocellular carcinoma in vivo and in vitroMethods After the expression of NCoA6 was knocked down with siRNA or lentivirus,the cell proliferation was confirmed by cell counting kit-8(CCK-8),clone formation,and EdU assay,and the cell apoptosis and cell-cycle were assessed by flow cytometry,while cell migration and invasion were investigated in the 24-transwell cell migration and collagen-based cell invasion system in vitro.Meanwhile,Western blot was used to tested the related proteins.In addition,the xenografted hepatic tumor model on nude mice was used to validate the tumor suppressing effect in vivo.Results Knockdown of NCoA6 in HCC cell lines of Huh7 and HCC-LM3 attenuated proliferation and influenced apoptosis and cell-cycle with alteration of the related proteins,but knockdown has no effect on cell migration and invasion as well as epithelial-mesenchymal transition(EMT)proteinsConclusions Knockdown of NCoA6 in Huh7 and HCC-LM3 influenced proliferation,apoptosis and cell cycle,while has no effect on cancer cell migration and invasion.NCoA6 functions as a candidate oncogenes in HCCSection three:NCoA6 promotes the proliferation of hepatocellular carcinoma through the PI3K/Akt/mTOR and the NF-?B signaling pathways.Methods The cDNA microarray was used to screen the downstream target genes regulated by NCoA6.After verifying the reliability of cDNA microarray results by qRCR,GO and KEGG enrichment analysis was carried out to determine the main biological functions and pathways of different genes.According to the results of enrichment analysis and the literature,the possible regulatory signaling pathways were selected,and the related proteins of these pathways were detected by Western blot.Results Based on the results of enrichment analysis and literature,we found that NCoA6 may regulate the occurrence and development of hepatocellular carcinoma through PI3K/Akt/mTOR and NF-?B signaling pathways.Western blot analysis showed that knockdown of NCoA6 could inhibit the activity of these two pathwaysConclusions NCoA6 promotes cell cycle progression and inhibits apoptosis of hepatocellular carcinoma cells through PI3K/Akt/mTOR and NF-?B signaling pathways.Part two:laparoscopic right hepatectomy for hepatocellular carcinoma:a propensity score matching analysis of outcomes compared with conventional open surgeryMethods Data of patients who underwent right hepatectomy for HCC from May 2007 to February 2019 in Hospital were obtained.Baseline characteristics,postoperative recovery,and survival outcomes were compared.One-to-one propensity score matching(PSM)was used to minimize selection biases by balancing factors including age,sex,preoperative therapy,and tumor size and pattern.Results The original cohort included 131 patients(LRH,51 patients;ORH,80 patients).Of the 51 patients who underwent LRH,8 patients(15.7%)required conversion to laparotomy.The overall morbidity was 19.6%,and no mortality in LRH was noted.After PSM,the operative time was comparable between two group(248.4 ±88.7 vs.231.6 ± 44.4 min,p=0.26),whereas less intraoperative blood loss[300(100-1200)vs.500(200-2000)mL,p<0.01]was observed in LRH group.LRH showed up a trend of less overall morbidity without statistically significant(18.2 vs.29.5%,p=0.21).Moreover,the 3-year overall and disease-free survival did not differ significantly between the groups during a median follow-up of 17(2-68)months for the LRH group and 15(3-103)months for the ORH group.Conclusions LRH can be performed as safely and effectively as ORH for HCC in regard to both surgical and oncological outcomes.LRH holds the benefit in less intraoperative blood loss and appears to achieve less postoperative morbidity and short hospital stay.LRH could serve as a promising alternative to ORH in selected individuals However,more methodologically high-quality comparative studies are required to adequately evaluate the status of LRH for HCC.