【Abstract】Objective To investigate the expression of inducible nitric oxide synthase (iNOS) and p53 protein in hepatocellular carcinoma (HCC) and their relationship with angiogenesis. Methods Immunohistochemical method and image analysis technique were used to detect the expression of iNOS and p53 protein in tumor tissue sections of 59 HCC patients. Microvessel density (MVD) was counted by immunohistochemical staining with anti-CD34 antibody.Results ①The expression rates of iNOS and p53 were 81.4%(48/59), 64.4%(38/59) in HCC patients, respectively. The expression intensities of iNOS and p53 were 5 635±1 287, 3 352±873 in HCC patients, respectively. ②MVD was 32.5±2.73 in the tumor tissue of HCC patients. ③The expression of iNOS was correlated with the expression of p53 and MVD in HCC patients (P＜0.05); The expression of p53 was also correlated with the MVD in HCC patients (P＜0.05). Conclusion iNOS and p53 are highly expressed in HCC and may play a key role in angiogenesis of HCC.
Objective To investigate the expressions of CD90, IGF1R, and hTERT protein in hepatocellular carcinoma, and the correlations of each other in the development of carcinoma. Methods The expressions of CD90, IGF1R, and hTERT protein in hepatocellular carcinoma were detected by S-P immunohistochemical staining, 20 cases of normal liver tissues were collected as contrast, and to compare the relations between expression and prognosis or survival rate. Results The positive rate of CD90, IGF1R, and hTERT protein in hepatocellular carcinoma group were obviously higher than that in contrast group（P＜0.05）, which was 63.9% vs. 0, 52.8% vs.5.0%, and 47.2% vs.0, respectively. The positive rate of CD90, IGF1R, and hTERT protein were higher in UICC Ⅲ-Ⅳ stage group than that in UICC stage Ⅰ-Ⅱ group（P＜0.05）, which was 79.2% vs.33.3%, 70.8% vs.16.7%, and 62.5% vs.16.7%, respectively. There was a statistically significant positive correlation observed between the expressions of CD90 and IGF1R protein （Kendall’s tau-b=0.563 1, P＜0.05）, so it was with CD90 and hTERT protein （Kendall’s tau-b=0.363 6, P＜0.05）. The survival rates of positive expressions of CD90, IGF1R, and hTERT protein were lower than negative expressions of CD90, IGF1R, and hTERT（P＜0.05）, which was 21.7% vs.50.0%, 17.6% vs.43.8%, and 20.0% vs.38.9%, respectively. Conclusions The expressions of CD90, IGF1R, and hTERT may have correlations with the progress of HCC, and may serve as a marker for HCC prognosis potentially.
Objective To investigate the association of the expression of CD15 mRNA with the invasion and prognosis of hepatocellular carcinoma (HCC) and the expression of nm23H1 mRNA. Methods In situ hybridization and immunohistochemistry methods were used to detect the expression of CD15 mRNA and protein nm23H1 mRNA in HCC.Results In 99 cases of HCC, the positive rate of CD15 mRNA,its protein and nm23H1 mRNA were 38.4%, 36.4% and 76.8%, respectively. The expression of CD15 mRNA was consistent with its protein and negatively correlated with the expression of nm23H1 mRNA. The expression of CD15 mRNA and its protein, nm23H1 mRNA were associated with the invasiveness and metastasis of HCC and the prognosis of HCC patients. Conclusion The detection of CD15 expression could be a new pathological biology index to judge the metastasis and prognosis of HCC.
【Abstract】ObjectiveTo construct a recombinant adeno-associated virus(rAAV2) vectors carrying the combined transcriptional regulatory sequences of α-fetoprotein enhancer and albumin promoter for the purpose of targeted gene therapy for hepatocellular carcinoma (HCC). MethodsThe fragment of combined transcriptional regulatory sequences of α-fetoprotein enhancer and albumin promoter was amplified through polymerase chain reaction (PCR) and cloned into the promoter site of pAAV-IRES-hrGFP instead of the CMV promotor in AAV Helper-Free System to construct the rAAV2 expression plasmid pAAV-IRES-hrGFP-EP. Then the packaging cell lines (HEK 293 cell) was co-transfected with the pAAV-IRES-hrGFP-EP together with the control plasmid pAAV-RC and pHelper in AAV Helper-Free System by means of lipofectamine.The recombinant adenoassociated virus vector(rAAV2-EP) carrying the combined transcriptional regulatory sequences of α-fetoprotein enhancer and albumin promoter was packaged and amplified in the HEK 293 cell. Then the viral titer was checked by GFP. ResultsThe recombinant adeno-associated virus vector(rAAV2-EP) carrying the combined transcriptional regulatory sequences of α-fetoprotein enhancer and albumin promoter was constructed successfully, the b green fluorescence was observed in HEK 293 cells under fluorescence microscope. The viral titer was 1.2×105. ConclusionConstruction of the recombinant adeno-associated virus vector rAAV2-EP driven by the combined transcriptional regulatory sequences of α-fetoprotein enhancer and albumin promoter would provide a sound basis and improved vector for targeted gene therapy for HCC.
Objective To summarize the papers about the research status and prospects of ferroptosis in hepatocellular carcinoma (HCC) and its drug resistance in recent years in order to provide directions and ideas for the treatment of HCC. Method The relevant literatures at home and abroad in recent years about ferroptosis in HCC and its drug resistance were reviewed. Results The mechanism of ferroptosis in the development and drug resistance of HCC was complicated, involving multiple protein and molecular pathways. Ferroptosis played an important role in improving chemotherapy and sorafenib resistance, and it had a broad application prospect in HCC. Conclusions The molecular mechanism of ferroptosis in HCC and its drug resistance has not been fully elucidated. Further research on the mechanism of ferroptosis in HCC may provide new molecular therapeutic targets for HCC. Ferroptosis has a broad application prospect in the treatment of HCC.
ObjectiveTo analyze the correlations between the immune function and inflammatory factors levels of patients with hepatocellular carcinoma (HCC) and the results of in vitro high-throughput drug sensitivity, and to provide a reference for personalized drug selection for patients with HCC. MethodsThe patients with HCC who met the inclusion criteria from December 2019 to June 2021 in the First Affiliated Hospital of Chongqing Medical University were included. The HCC cells were used to perform in vitro high-throughput drug sensitivity screening, the result was classified into sensitive and insensitive. The correlations between drug sensitivity results and immune function and inflammatory factors levels of corresponding patients were analyzed, and the relation between these indexes (P<0.05) and drug sensitivity of HCC cells to drugs or combination regimen of drugs was further analyzed by univariate logistic regression. ResultsA total of 74 patients with HCC were included in this study. The results showed that the level of interleukin-6 was negatively correlated with sorafenib, caffezomib, gemcitabine, oxaliplatin + epirubicin + irinotecan + 5-fluorouracil, oxaliplatin + irinotecan + epirubicin, and oxaliplatin + epirubicin regimens on the inhibition rates of HCC in vitro (P<0.05), and positively correlated with bortezomib (P<0.05). However, the level of interleukin-6 was not related to the sensitivity of HCC cells to these single drugs or combined regimens (P>0.05). Meanwhile it was found that tumor necrosis factor (TNF)-α was negatively correlated with cabotinib, apatinib, caffezomib, and epirubicin on the inhibition rates of HCC in vitro (P<0.05), and positively correlated with epirubicin (P<0.05). But only it was found that tumor necrosis factor-α level was related to the sensitivity of HCC cells to epirubicin (P<0.05). ConclusionsTumor necrosis factor-α level in peripheral blood of patients with HCC has a certain relation with epirubicin on inhibition rate of HCC in vitro and it might have a certain value in predicting sensitivity of HCC cells to epirubicin. Meanwhile, although it is found that level of IL-6 is related to sorafenib, caffezomib, gemcitabine, or including combination regiems including oxaliplatin and epirubicin on inhibition rates of HCC in vitro, their value is not found in predicting sensitivity of HCC cells to these single drugs or combined regimens.
ObjectiveTo summarize the comprehensive multidisciplinary team (MDT) treatment and holistic scientific management approach in a case of hepatocellular carcinoma (HCC) in S7+S9 segments with prostatic metastasis. MethodsA retrospective analysis was conducted on the clinical data and follow-up outcomes of a patient with HCC in S7+S9 segments who developed prostatic metastasis during treatment, admitted to the Tenth Affiliated Hospital of Southern Medical University (Dongguan People’s Hospital of Guangdong Province). ResultsDue to the complexity of the patient’s condition, an MDT discussion was held upon initial admission. It was concluded that the HCC diagnosis was clear, with lesions confined to the liver (S7+S9 segments) and a tumor diameter less than 3 cm, making surgical resection or ablation therapy the preferred options. However, the patient declined liver transplantation and surgical resection. Therefore, CT-guided microwave ablation (MWA) was performed on the primary HCC lesions in segments S7 and S9b. Prior to subsequent treatments for recurrent disease, MDT discussions were held again, and treatments were tailored to the discussion outcomes while respecting the patient’s wishes. Over time, the patient underwent CT-guided liver puncture MWA, re-ablation for recurrent tumors, transarterial chemoembolization (TACE), stereotactic body radiation therapy (SBRT), targeted therapy, and immunotherapy. Following this comprehensive MDT treatment plan, the patient had survived for over 78 months, with no evidence of active tumor lesions in the liver, prostate, or other parts of the body. Alpha-fetoprotein levels and liver function remained normal, and the patient’s quality of life was good. ConclusionComprehensive MDT treatment incorporates various technologies and approaches, along with holistic scientific management, can yield favorable outcomes for patients with complex and challenging HCC.
Objective To investigate the cell growth inhibition and apoptosis induced by rapamycin on human hepatocellular carcinoma Bel-7402 cells and to study the role of mitochondrium membrane potential in the process of apoptosis. Methods Bel-7402 cells in vitro were given 5, 10, 20, 30, 40 and 50 nmol/L different concentrations of rapamycin, and the cell growth inhibiting ratio of Bel-7402 was assessed by MTT assay. The changes of morphology of Bel-7402 were observed by Hoechst 33258 staining and flow cytometry (FCM), respectively; The cell mitochondrial membrane potential was detected by using JC-1 staining method. Results Rapamycin could inhibit the growth of Bel-7402 cells significantly by inducing apoptosis, and the growth suppression and the cell apoptosis both presented time-effect relationship and were also dose-dependent. The rates of inhibiting and cell apoptosis after 72 h exposure to 50 nmol/L rapamycin were significantly higher that those of other groups (P＜0.01). Typical morphological changes of cell apoptosis were observed very clearly after the Bel-7402 cells had been exposed to rapamycin for 48 hours using Hoechst 33258 staining method, and it was also observed that the mitochondrial membrane potential decreased when apoptosis occured (P＜0.01). Conclusion Rapamycin could inhibit the growth of Bel-7402 cells by inducing cell apoptosis, and the descent of mitochondrial membrane potential may play an important role in the process of cell apoptosis.
ObjectiveTo investigate the effect of the round ligament fissure approach in re-hepatectomy.MethodsA total of 40 patients with recurrence of hepatocellular carcinoma (HCC) who underwent re-hepatectomy in the Department of Hepatopancreatobiliary Surgery of Leshan People’s Hospital from June 2017 to August 2020 were collected and divided into two groups according to different surgical approaches: study group (transhepatic round ligament fissure approach) and control group (conventional surgical approach), 20 cases in each group. The perioperative general indicators, peripheral blood laboratory indicators, and complications of the two groups were compared.ResultsCompared with the control group, the operation time, postoperative drainage tube removal time, and postoperative hospital stay of study group were shortened, and intraoperative blood loss was reduced (P<0.05). Compared with preoperatively in the same group, postoperative TBIL and ALT levels of the two groups decreased, and HGF levels increased (P<0.05). There was no significant difference in the levels of TBIL, ALT, and HGF between the two groups before surgery (P>0.05); at 1 month after surgery, there was no significant difference in the levels of TBIL and ALT between the two groups (P>0.05), but the HGF level of the study group was higher than that of the control group at1 month after operation, the difference was statistically significant (P<0.05). The changes before and after operation of TBIL and ALT were similar between the two groups (P>0.05), but the rising value of HGF in the study group was higher than that of the control group (P<0.001). There was no death in the two groups during the perioperative period, and the total postoperative complications were not statistically different (P=0.677). There was no statistically significant difference in the postoperative follow-up results between the two groups in recurrence, metastasis, and death (P>0.05).ConclusionRe-hepatectomy through the round ligament fissure approach can reduce the amount of intraoperative blood loss, shorten the operation time, and reduce the damage to the residual liver, which has high safety.