Objective To explore the effects of overexpression of human tissue inhibitors of metalloproteinase-1 (hTIMP-1) on proliferation of human liver cancer cell line HepG2 in vitro. Methods A recombinant adenoviral vector containing full-length cDNA of hTIMP-1 was generated and transfected into HepG2. The viral titer was checked by measuring GFP, and the expression of hTIMP-1 in vitro was detected by the techniques of Western blot and semi-quantitative RT-PCR. The ultrastructure was observed by transmission electron microscope and the effects of overexpression of hTIMP-1 on proliferation of HepG2 in vitro was analyzed by MTT assay and growth curve. Results The resultant AdhTIMP-1 was successfully constructed and the expression of hTIMP-1 was detected by Western blot and RT-PCR. The growth and proliferation of HepG2, which had been transfected with AdhTIMP-1, was significantly inhibited. Conclusion The proliferation of HepG2 was markedly inhibited by recombinant adenovirus-mediated overexpression of hTIMP-1, which may pave the way for further application in liver gene therapy.
Objective To identify genes associated with hepatocellular carcinoma (HCC) as candidate diagnostic markers in a genome-wide scale. Methods The gene expression profiles of 40 pairs of HCC tumor tissue and peripheral non-tumorous liver tissue were analyzed by using gene chip technology.The gene chips were fabricated at the National Cancer Institute (NCI). Each gene chip contained 9 180 genes. The fluorescent targets were prepared by a direct labeling approach using two kinds of fluorescences as following: 100 μg of total RNA from non-cancerous liver tissue was labeled with Cy3-dUTP and 200 μg of total RNA from HCC was labeled with Cy5-dUTP. The targets were mixed together and hybridized with genes on the gene chips. Unsupervised hierarchical clustering analysis was done by CLUSTER and TREEVIEW software using median centered correlation and complete linkage. Results A total of 10 genes were found up-regulated in over 80% of primary tumors comparing with that of their corresponding non-tumorous liver tissues at a two-fold filter with an unsupervised hierarchical clustering algorithm, including protocadherin-alpha 9, ESTs, Homo sapiens cDNA FLJ, KPNA2, RPS20, SNRPE, CDKN2A, UBD, MDK and ANXA2.Conclusion These genes are supposed to be candidates for the diagnosis of HCC. Further investigation of these genes in a large scale of patients with HCC and patients with non-malignant hepatic diseases will be needed to disclose whether they could be used clinically as novel diagnostic tumor markers for HCC.
Objective To explore the value of laparoscopic hepatectomy for small hepatocellular carcinoma (HCC) of non-peripheral type. Methods The clinical data of 34 patients with small HCC of non-peripheral type underwent laparoscopic liver resection from March 2008 to April 2011 in our hospital were analyzed retrospectively. Results Thirty-two patients received successful total laparoscopic hepatectomy without blockage of liver blood flow，and 2 were converted to open surgery. The operative time was （162±65） min （100-220 min） and the blood loss was （295±166） ml （100-750 ml）. There were postoperative complications in 4 patients， included cross-section bleeding in 2 cases and ascites in 2 cases. There were no complications such as biliary fistula， infection， carbon dioxide gas embolism， and so on. The mortality of perioperative period was 0. The postoperative hospital stay was （6±2） d （4-9 d）. The follow-up time was （23±7） months （5-42 months）. Thirteen patients developed intrahepatic tumor recurrence during follow-up. The overall and recurrence-free survival rate one year after operation was 90.6% （29/32）and 75.0% （24/32）， respectively. Conclusions Laparoscopic hepatectomy is a safe， feasible， and minimal invasive approach for small HCC of non-peripheral type，and it can be considered as a alternative treatment of HCC.
Objective To investigate the effects of adenovirus-mediated melanoma differentiation-associated gene-7 (mda-7)/IL-24 and/or adriamycin (ADM) on transplanted human hepatoma in nude mice and to explore a new way for hepatoma gene therapy combined with chemotherapy. Methods The recombinant adenovirus vector carrying Ad.mda-7 was constructed; Ad.mda-7 and/or ADM were injected into the tumor-bearing mice. Their effects on the growth of the tumor and the survival time of the mice were observed. The expressions of VEGF and TGF-β1 were detected by an immunohistochemistry method. Results Ad.mda-7 was constructed and expressed in vivo successfully. Compared with other three groups 〔control group (43.4±1.67) d, ADM group (64.2±4.14) d, Ad.mda-7 group (61.4±1.67) d〕, the mice treated with Ad.mda-7 combined with ADM had longer average survival time 〔(83.8±4.82) d, P＜0.01〕; the average size of tumor treated with Ad.mda-7 combined with ADM diminished significantly compared with that treated with ADM or Ad.mda-7 separately (P＜0.01). VEGF and TGF-β1 expressions of Ad.mda-7 group were (56.2±7.7)%, (35.2±4.5)%, respectively, and were lower than those in ADM group (VEGF: P＜0.05; TGF-β1: P＜0.01). VEGF expression of Ad.mda-7+ADM group was (37.3±5.0)%, and was significantly lower than that in other three groups (P＜0.01). TGF-β1 expression of Ad.mda-7+ADM group was (31.2±3.1)% and significantly lower than that in control group and ADM group (P＜0.01), however, there was no significant difference compared with Ad.mda-7 group (Pgt;0.05). Conclusion Ad.mda-7 combined with ADM has b antitumor potency and synergistic effects and suppresses the growth of human HCC xenograft in nude mice, possibly by inducing the apoptosis of hepatoma cell lines and suppressing tumor angiogenesis.
ObjectiveTo study the relationship between expression of p27KIP1 and progression of hepatocellular carcinoma(HCC).MethodsThe expression of p27KIP1 in 52 cases of HCC was detected by immunohistochemistry of strept avidinbiotin complex and mRNA in situ hybridization.ResultsThe positive cells of p27KIP1 protein were diffused in HCC.The positive signal was localized in nuclei.The labeling index (LI) of p27KIP1 protein was significantly higher in tumorsurrounding tissues than that in tumor tissues. p27KIP1 protein LI showed a positive correlation with the differentiation grade of HCC.The better differentiation of cancer cells, the higher LI of p27KIP1 protein (P＜0.01).The positive cells of p27KIP1 mRNA were also diffused in HCC.The positive signal was localized in nuclei and cytoplasm. As to the expression of p27KIP1 at the mRNA level,there was no significant correlation with tumorsurrounding tissues and stages of HCC.Conclusionp27KIP1 protein is associated with progression and differentiation grade of hepatocellular carcinoma.
ObjectiveTo explore the clinical, imaging, and pathological features of patients with synchronous double primary hepatocellular carcinoma and intrahepatic cholangiocarcinoma (sdpHCC-ICC), to enhance our understanding of the disease and reduce the rate of misdiagnosis and missed diagnosis.MethodsThe clinical, imaging, and pathological data of patients who were histologically confirmed as sdpHCC-ICC in West China Hospital of Sichuan University between January 1st 2014 and December 31st 2018 were studied retrospectively.ResultsA total of 11 patients with sdpHCC-ICC were screened for the study, of which 10 were male and 1 was female. The median age of patients was 55.6 years (ranged from 47 to 73 years). Eight patients were chronically infected with hepatitis B virus. Both increased alpha-fetoprotein and carbohydrate antigen 19-9 were observed in 8 patients. Contrast enhanced CT was performed in 8 cases, color doppler ultrasound in 4 cases, enhanced MRI in 3 cases, and contrast-enhanced ultrasound in 1 case. Among them, one solitary lesion was found in 2 patients, and two or more lesions were observed in 9 patients. Most of the patients had typical imaging performance of hepatocellular carcinoma (HCC): 8 patients showed strong enhancement of HCC during the hepatic arterial phase and progressive hyper-attenuation on venous and delayed phases, 1 patient showed peripheral rim enhancement in the arterial phase of intrahepatic cholangiocarcinoma (ICC) in another lesion could be observed at the same time. None of the 11 patients with sdpHCC-ICC was diagnosed accurately before operation. All patients underwent surgical treatment. HCC lesions were distributed in all parts of the liver, while ICC lesions were located in the right lobe of the liver in 10 cases. The median diameter of HCC and ICC was 3.5 cm and 2.1 cm, respectively. All of them were confirmed by hematoxylin-eosin staining and immunohistochemistry.ConclusionsThe clinical characteristics of sdpHCC-ICC are usually atypical. It is difficult to make an accurate preoperative diagnosis. Tumor markers may be valuable to the diagnosis of sdpHCC-ICC. The definite diagnosis of sdpHCC-ICC depends on pathological examination.
ObjectiveTo explore the relationship between the expressions of fibronectin (FN) and phosphatase and tensin homology deleted on ehromosome ten (PTEN) in hepatocellular carcinoma (HCC) tissues and the clinical pathological features. MethodsThe expressions of FN and PTEN were detected by using Western blot and immunohistochemistry respectively in 83 HCC tissues and para-carcinoma tissues, 47 hepatic cirrhosis tissues and 11 normal hepatic tissues. The correlations between the expressions of FN and PTEN and the clinicopathologic features of HCC patients were analyzed. ResultsThe positive expression rate of FN protein in HCC tissues was significantly higher than those in para-carcinoma tissue, normal hepatic tissue, and liver cirrhosis tissues (P<0.05); meanwhile the expression of PTEN was opposite (P<0.05). The positive expression rate of FN protein in para-carcinoma tissues was also obviously higher than that liver cirrhosis tissues and normal hepatic tissues (P<0.05), meanwhile the expression of PTEN was opposite (P<0.05). The positive expression rate of FN protein was higher in HCC tissues with cancer embolus, lymphatic metastasis, positive AFP, and multiple tumor (P<0.05), but there were no statistically significant differences in FN protein expression, gender, age, HBsAg, degree of tumor differentiation, and size of tumor (P>0.05). The positive expression rate of PTEN was lower in HCC tissues with high-medium differentiation, cancer embolus, positive AFP, lymphatic metastasis, and tumor diameter ≥2 cm (P<0.05), there were no statistically significant differences in PTEN expression, gender, age, HBsAg, and the number of tumor (P>0.05). ConclusionsThe abnormal expressions of FN and PTEN in HCC tissues which may play a role in promoting or inhibiting occurrence, development, invasion, and metastasis of HCC. The abnormal expressions of both can be used as molecular biological markers for the malignant degree, invasion, and metastasis of HCC.
ObjectiveTo establish multidrugresistance cell substrain of human hepatocellular carcinoma and to investigate its characteristics.MethodsSMMC7721 cell strain was cultured in Adriamycin(ADM). The multidrugresistance cell substrain SMMC7721/ADM was harvested after a long period of culture by gradually increasing the concentration of ADM and its characteristics were investigated. Results①The drug resistance of SMMC7721/ADM to ADM increased by 33.3 times, to Vincristine 16.8 times, to Diamminedichloroplatinum 2.8 times. ②The drug resistance cell substrain had almost the same growth velocity as its parental generation. The doubling time was 32.0 hours and 30.5 hours respectively. They had the analogous growth curves. ③The obvious difference between the drug resistance cell substrain and its parental generation was that the former’s microvilli became thick, short and scattered by scanning and transmitting electron microscopy. ④The multidrug resistance cell substrain kept the characteristics of hepatocellular carcinoma, it could be transplanted into the subcutaneous tissue of nude mice. ⑤The drug resistance of the cell substrain reduced to 28.0% and 9.2%after removal of the drug for 1 month and 2 months respectively, its drug resistance could remain stable (35.4 times) after 2 months of culture in ADM (0.04 μg/ml).ConclusionThe SMMC7721/ADM cell substrain has the stable fundamental characteristics of a drug resistance cell strain.
Objective To study the relationship of hepatitis B virus （HBV） to spontaneous rupture of hepatocellular carcinoma （HCC-SR） and its mechanism. Method The related literatures about theory of HCC-SR were consulted and reviewed. Results The injury of small arteries was usually followed in patients with HCC-SR， which was related to vascular autoimmune injury caused by the HBV infection. The small arteries in which immune complex deposited were readily injured， as a result HCC-SR happened while vascular load increased. Conclusion The HBV infection resulted in vascular autoimmune injury maybe a important factor in the pathogenesis of HCC-SR.
Two hundred and thirty patients with solid hepatic space-occupying lesions (SHSOL), on whom hepatic resection was performed in Zhongshan hospital, were analyzed. We found that liver cirrhosis could be a diagnostic marker of hepatocellular carcinoma in patients with SHSOL, for which the sensitivity being 85.2%, the specificity 96.3%, and the positive predictive value 98.7%.