The Chinese human hepatocellular carcinoma cell SMMC7721 has been analysed by flow cytometry, Southern blotting and Western blotting. The results indicated that SMMC7721 cell is a hypoploid cell with a 0.81 DNA index, and that SMMC7721 cell has internal deletion in the 5'-end of its Rb gene and has no Rb gene product (Rb protein). The normal Rb cDNA has been inserted into a retrovirus vector DOL and introduced into SMMC7721 cells by electrporation transfection technique.About 75% of transfected SMMC7721 cells have been killed by Rb gene product. The remain 25% cells are alive as exogenous Rb gene has been mutationally inactivated. （Chin J Ocul Fundus Dis,1994,10:21-24）
ObjectiveTo understand the interplay between tumor-associated macrophages (TAMs) and T lymphocytes, as well as the effect on the pathogenesis of hepatocellular carcinoma (HCC) again, so that providing new ideas and methods for the immunotherapy of HCC.MethodSearched the literatures about the interplay between TAMs and T lymphocytes in HCC to analyze and summarize the relationship between TAMs and T lymphocytes in HCC.ResultsWhile TAMs and T lymphocytes themselves regulate the process of tumorigenesis and development, they also had a mutual regulatory mechanism to further promote the development of HCC.ConclusionsThere is an interaction between TAMs and T lymphocytes, and this interaction forms a vicious circle to a large extent and promotes the development of HCC. Recognizing and making rational use of this interaction can provide new ideas and methods for the future immunotherapy of HCC.
For an advanced elucidation of mechanisms of nm23-H1 suppressive effects on metastasis of primary hepatocellular carcinoma (HCC), it is necessary to investigate the correlation between nm23-H1 expression and relative factors involved in the HCC invasion. In present report, full-length cDNA of nm23-H1 was subcloned into pBKCMV vector and transfected into HCC cell line to observe its effects on invasion, cytosolic free Ca2+ and Nras mRNA expression. The results showed that lower expression of N-ras and higher cytosolic free Ca2+ in transfected cell line were detected, while the potential of invasion was depressed. It suggests that the suppressive effects on HCC metastasis might interact with intracellular signal transduction which is essential for stimulating cell invasion.
Objective To summarize the research progress of magnetic resonance imaging (MRI) in evaluating microvascular invasion (MVI) of hepatocellular carcinoma (HCC) in order to provide information and evidence for therapy of HCC. Methods Papers published from May 1950 to May 2017, were retrieved in PubMed, OVID, CNKI database using the keywords on hepatocellular carcinoma, microvascular invasion, and magnetic resonance imaging. Sixty-seven papers were retrieved in English literatures and 13 in Chinese literatures. Criteria of paper adoption: ① the imaging method was MRI; ② the assessment content was MVI of HCC; ③ the golden standard was postoperative pathologic diagnosis. fifty-four papers were finally analyzed and reviewed. Results Currently there were various ways to evaluating the MVI of HCC using MRI, including morphology, texture analysis, diffusion-weighted imaging, dynamic-enhanced MRI, fat assessment, hepatocellular function and comprehensive evaluation. Conclusions Various methods perform differently in evaluating MVI. The use of multiparametric MRI techniques offers the potential for comprehensive assessment of MVI of HCC.
ObjectiveTo summarize the research progress of hepatocellular carcinoma (HCC) based on tumor microenvironment immunophenotyping.MethodThe related literatures of basic and clinical studies on HCC immunophenotyping in the recent years were reviewed.ResultsHCC could be divided into different immunophenotypes based on tumor microenvironment, and it showed different immune molecular characteristics, immune cell infiltration characteristics, and anti-tumor ability. At the same time, the HCC immunophenotype was significantly associated with patients’ survival and had been proved to be able to better evaluate the prognosis of HCC patients. According to the relevant molecular characteristics in the HCC immune microenvironment, it could provide guidance for the drug regimen of immunotherapy.ConclusionHCC immunophenotyping is still in the early stage of research, and its clinical application value has been preliminarily shown for the evaluation of patients’ prognosis and immunotherapy decision-making, which is a new idea of individualized treatment of HCC in the future.
ObjectiveTo investigate the feasibility of associating liver partition and portal vein ligation for staged hepatectomy (ALPPS) combined with mixed reality holographic imaging in treating the massive hepatocellular carcinoma.MethodThe clinicopathologic data of 1 patient with massive hepatocellular carcinoma underwent the ALPPS combined with mixed reality holographic imaging in the Guangdong Second Traditional Chinese Medicine Hospital on January 2019 were retrospectively analyzed.ResultsA 58-year-old female patient, the preoperative CT scan and enhanced scan of the abdomen revealed a 12.0 cm×10.5 cm×17.0 cm mass in the right lobe of the liver, with preoperative Child-Pugh grade A liver function. The ALPPS was performed after the preoperative evaluation. The associating liver partition and portal vein ligation was performed and the mixed reality holographic imaging was used during the stage Ⅰ operation. On day 21 after the stage Ⅰ operation, the CT results of the volume of left lateral lobe of liver was approximately 57.64%. On day 24 after the stage Ⅰ operation, the patient underwent the stage Ⅱ operation (radical hepatectomy). The patient received the chemotherapy after the operation. The courses of those operations were successful. And no severe complications occurred after the operation. The postoperative pathological results showed a moderately differentiated hepatocellular carcinoma with multiple foci and a maximum diameter of 13 cm. Six months after the last operation, there was no new intrahepatic metastasis and other metastases were found.ConclusionsALPPS combined with mixed reality holographic imaging is effective in treatment of massive hepatocellular carcinoma. Mixed reality holographic imaging could show anatomical details with three dimensions, it is beneficial for estimating future liver reserve before operation and locating during operation, and there is also a positive significance of postoperative recovery.
ObjectiveTo screen long non-coding RNAs (lncRNAs) relevant to programmed cell death (PCD) and construct a nomogram model predicting prognosis of hepatocellular carcinoma (HCC). MethodsThe HCC patients selected from The Cancer Genome Atlas (TCGA) were randomly divided into training set and validation set according to 1∶1 sampling. The lncRNAs relevant to PCD were screened by Pearson correlation analysis, and which associated with overall survival in the training set were screened by univariate Cox proportional hazards regression (abbreviation as “Cox regression”), and then multivariate Cox regression was further used to analyze the prognostic risk factors of HCC patients, and the risk score function model was constructed. According to the median risk score of HCC patients in the training set, the HCC patients in each set were assigned into a high-risk and low-risk, and then the Kaplan-Meier method was used to draw the overall survival curve, and the log-rank test was used to compare the survival between the HCC patients with high-risk and low-risk. At the same time, the area under receiver operating characteristic curve (AUC) was used to evaluate the value of the risk score function model in predicting the 1-, 3-, and 5-year overall survival rates of HCC patients in the training set, validation set, and integral set. Then the nomogram was constructed based on the risk score function model and factors validated in clinic, and its predictive ability for the prognosis of HCC patients was evaluated. ResultsA total of 374 patients with HCC were downloaded from the TCGA, of which 342 had complete clinicopathologic data, including 171 in the training set and 171 in the validation set. Finally, 8 lncRNAs genes relevant to prognosis (AC099850.3, LINC00942, AC040970.1, AC022613.1, AC009403.1, AL355974.2, AC015908.3, AC009283.1) were screened out, and the prognostic risk score function model was established as follows: prognostic risk score=exp1×β1+exp2×β2...+expi×βi (expi was the expression level of target lncRNA, βi was the coefficient of multivariate Cox regression analysis of target lncRNA). According to this prognostic risk score function model, the median risk score was 0.89 in the training set. The patients with low-risk and high-risk were 86 and 85, 86 and 85, 172 and 170 in the training set, validation set, and integral set, respectively. The overall survival curves of HCC patients with low-risk drawn by Kaplan-Meier method were better than those of the HCC patients with high-risk in the training set, validation set, and integral set (P<0.001). The AUCs of the prognostic risk score function model for predicting the 1-, 3-, and 5-year overall survival rates in the training set were 0.814, 0.768, and 0.811, respectively, in the validation set were 0.799, 0.684, and 0.748, respectively, and in the integral set were 0.807, 0.732, and 0.784, respectively. The multivariate Cox regression analysis showed that the prognostic risk score function model was a risk factor affecting the overall survival of patients with HCC [<0.89 points as a reference, RR=1.217, 95%CI (1.151, 1.286), P<0.001]. The AUC (95%CI) of the prognostic risk score function model for predicting the overall survival rate of HCC patients was 0.822 (0.796, 0.873). The AUCs of the nomogram constructed by the prognostic risk score function model in combination with clinicopathologic factors to predict the 1-, 3-, and 5-year overall survival rates were 0.843, 0.839, and 0.834. The calibration curves of the nomogram of 1-, 3-, and 5-year overall survival rates in the training set were close to ideal curve, suggesting that the predicted overall survival rate by the nomogram was more consistent with the actual overall survival rate. ConclusionThe prognostic risk score function model constructed by the lncRNAs relevant to PCD in this study may be a potential marker of prognosis of the patients with HCC, and the nomogram constructed by this model is more effective in predicting the prognosis (overall survival) of patients with HCC.
Objective To explore the feasibility of mean temporal phase images calculated from perfusion CT datasets by using CT perfusion (CTP) of liver on the third-generation dual-source CT. Methods Twenty-two consecutive patients with suspected hepatocellular carcinoma were enrolled, we retrospectively compared objective and subjective image quality, leson detectability, and radiation dose between mean temporal arterial (mTA) and mean temporal portal venous (mTPV) images which calculated from perfusion CT datasets with conventional enhanced arterial and portal venous datasets. Results ① Image quality: compared with the conventional enhancement image, the standard deviation (SD) values of CTP images on liver (arterial phase), portal vein (arterial phase), and liver (portal vein phase) were lower (P<0.05); the signal-to-noise ratio (SNR) values of CTP images on aorta (arterial phase), portal vein (arterial phase), aorta (portal vein phase), and portal vein (portal vein phase) were all higher (P<0.05), the contrast-to-noise ratio (CNR) value of CTP images on aorta (arterial phase) was higher (P<0.05). ② The subjective image quality: the subjective image quality scores of CTP images (mTA and mTPV images) were higher when compared to responding conventional enhanced arterial and portal venous datasets (P<0.05). ③ The diagnostic efficiency: the CTP images and conventional enhancement images showed all the lesions, but the diagnostic efficiency images of CTP images was better than the conventional enhancement images, both on lesions of blood supply and lack of blood supply (P<0.05). Conclusions The image quality of mTA and mTPV datasets calculated from CTP datasets are non-inferior when compared to conventional enhanced arterial and portal venous acquisitions in patients with suspected hepatic lesions. Thus, CTP could be used as a stand-alone imaging technique without additionally performed conventional arterial and portal venous CT acquisitions.
Objective To systematically evaluate the diagnostic efficacy of circulating tumor DNA (ctDNA) in hepatitis B viral hepatocellular carcinoma (HBV-HCC), and to study the clinical value of ctDNA. Methods The databases of PubMed, Embase, Web of Science, and Cochrane Library database were retrieved systematically from the establishment of the database to April 26, 2021. The characteristic information of literatures and the original data such as the sensitivity, specificity, and area under curve (AUC) of the receiver operating characteristic (ROC) curve were extracted. A meta-analysis was conducted by applying RevMan 5.3 and Stata 15.0 software. The combined sensitivity, combined specificity, positive likelihood ratio, negative likelihood ratio, and diagnostic odds ratio (OR) were calculated, ROC curve was plotted and the AUC was calculated, Deck’s funnel chart to assess publication bias, the Fagan diagram to test the diagnostic efficiency. Results Finally, 16 studies involving 3 744 patients were enrolled in this study, of which 1 852 were HBV-HCC patients, and 1 892 were HBV-infected patients without HCC. The meta-analysis results showed that ctDNA had a pooled sensitivity of 0.85 [95%CI (0.78, 0.90)], a specificity of 0.74 [95%CI (0.63, 0.83)], a diagnostic OR of 15.98 [95%CI (10.65, 23.99)], and the AUC of ROC was 0.87 [95%CI (0.84, 0.90)] in the diagnosis of HBV-HCC. The pooled sensitivity, specificity, diagnostic OR, and the AUC of ROC for ctDNA combined with AFP in the diagnosis of HBV-HCC were 0.86 [95%CI (0.80, 0.90)], 0.79 [95%CI (0.68, 0.87)], 22.69 [95%CI (13.64, 37.76)], and 0.90 [95%CI (0.87, 0.92)]. Meta-regression analysis found that the heterogeneity came from other non-covariate factors. The Fagan chart showed that while HBV-HCC was diagnosed by liquid biopsy-based on ctDNA, the probability of being diagnosed with hepatocellular carcinoma was 77%, if HBV-HCC was excluded, the probability of having the corresponding disease was 17%. Deek’s test showed no obvious publication bias (P>0.05). ConclusionsThe ctDNA can diagnose HBV-HCC with high sensitivity, specificity and accuracy, and can be used as a promising circulating biomarker in the early diagnosis of HBV-related HCC. The combination of ctDNA in serum and AFP is beneficial to improve the diagnostic accuracy of HBV-HCC.
【Abstract】ObjectiveTo construct an mdr1 expression vector and detect its expression in HepG2 cells in vitro. MethodsThe 4.5-kb mdr1 cDNA was obtained from the plasmid pHaMDR1 cloned into the PCIneo mammalian expression vector, which was later transferred into human hepatocarcinoma cell line HepG2 by liposome. Then the HepG2 cells resisting G418 were clustered and proliferated，and the specific fragment of mdr1 cDNA, mRNA and the Pgp in these HepG2 cells were detected by means of PCR, RT-PCR and FCM respectively. ResultsThe mdr1 expression vector was constructed successfully，and the stable multidrug resistance(MDR) hepatocarcinoma cell line （HepG2/mdr1） was developed as well. The outcome of PCR analysis showed that the specific fragment of mdr1 cDNA could be found in HepG2/mdr1 cells, but not in the nontransfection HepG2 cells. Furthermore,the content of the specific fragment of mdr1 mRNA and the expression of P-gp in HepG2/mdr1 cells were (59.7±7.9)% and (12.5±5.45)% respectively, the corresponding value in HepG2 cells were (16.9±3.2)% and (4.63±2.59)% respectively. The difference was statistically significant (P＜0.05). ConclusionIt is praticable to develop MDR hepatocarcinoma cell line by transferring mdr1 cDNA into HepG2 cells, which is useful in the research of MDR mechanism.