Investigating the cytotoxic effect of ibuprofen concentration in liver cancer cells(HepG2)and normal fibroblast(AGO)

Objective:Although many studies have reported that nonsteroidal anti-inflammatory drugs can have anticancer effects,the results are still challenging.The aim of this research is to Mechanism and effect of anti-inflammatory drugs in cancer treatment.Methods:In this laboratory study,cell lines were randomly divided into control group(no exposure to ibuprofen and groups exposed to ibuprofen concentrations of 10,1,0.1,and 0.001 mg/mL.The cytotoxic effect of ibuprofen was measured at 24 and 72 hours using the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide(MTT).Data were compared between groups using a one-way variance test.Results:The results showed that the viability rate of HepG2 cancer cells at concentrations of 1 and 10 mg/mL decreased significantly compared to the control group in 24 hours(P<0.0001).Also,the viability rate at concentrations of 1,10,0.1,and 0.001 mg/mL decreased significantly compared to the control group in 72 h(P<0.0001).Only the concentration of 10 mg/mL ibuprofen decreased the viability of normal cells compared to the control(P<0.05).Conclusion:Overall,the results of this research showed that different concentrations of ibuprofen had a cytotoxic effect on liver cancer cells,and except for the concentration of 10 mg/mL,the other concentrations did not have a cytotoxic effect on normal cells.

Dichloroacetate enhances the antitumor effect of pirarubicin via regulating the ROS-JNK signaling pathway in liver cancer cells

Aim:Liver cancer is one of the most common malignancies and has a high recurrence rate.However,current treatment strategies do not achieve satisfactory outcomes in the clinic.To explore a new strategy to enhance the effectiveness of chemotherapy in liver cancer,we investigated whether dichloroacetate(DCA)could enhance the sensitivity of liver cancer cells to pirarubicin(THP).Methods:Liver cancer cells were treated with DCA alone,THP alone,or DCA and THP combined.Cell viability was determined by the CCK-8 assay.Cell apoptosis was analyzed by flow cytometer.Reactive oxygen species(ROS)were detected using a CM-H2DCFDA fluorescence probe.Protein levels were identified by immunoblotting.Results:The results revealed that DCA significantly enhanced the antitumor effect of THP in liver cancer cells.Changes in morphology and adherence ability were observed,as well as decreased cell viability.The results of flow cytometry showed that the combination of THP and DCA significantly increased apoptosis of liver cancer cells.Moreover,compared with THP alone,combination treatment with DCA significantly increased THP-triggered ROS generation in liver cancer cells.The antioxidant N-acetyl-L-cysteine reversed the synergistic effect of DCA and THP on ROS generation,cell viability and apoptosis.Furthermore,phosphorylation of c-Jun N-terminal kinase(JNK)was significantly increased in the DCA and THP combination group.The effects of DCA and THP on cell viability and apoptosis were inhibited by the JNK inhibitor SP600125.Conclusion:The results obtained in the present study indicated that DCA enhanced the antitumor effect of THP in liver cancer cells via regulating the ROS-JNK signaling pathway.

Influence of adriamycin on changes in Nanog, Oct-4, Sox2, ARID1 and Wnt5b expression in liver cancer stem cells

AIM: To determine the influence of Adriamycin (ADM) on the changes in Nanog, Oct4, Sox2, as well as, in ARID1 and Wnt5b expression in liver cancer stem cells.

Regulators of liver cancer stem cells

Hepatocellular carcinoma(HCC)is a leading cause of cancer deaths.It is often detected at a stage when there are few therapeutic options.Liver cancer stem cells(LCSCs)are highly tumorigenic and resistant to chemotherapy and radiation therapy.Their presence in HCC is a major reason why HCC is difficult to treat.The development of LCSCs is regulated by a variety of factors.This review summarizes recent advances on the factors that regulate the development of LCSCs.Due to the importance of LCSCs in the development of HCC,a better understanding of how LCSCs are regulated will help to improve the treatments for HCC patients.

A STUDY OF THE FORMATION OF LIVER METASTASIS AND ITS MECHANISM USING THE INTRASPLENIC INOCULATION OF CANCER CELLS

In order to establish an animal model with hepatic metastasis intrasplenic inoculation of carcinoma cells from murine uterine cervical carcinoma (U14) was employed. Results showed a high incidence of hepatic metastasis could be obtained through the intrasplenic inoculation of 1 × 106 carcinoma cells. Removal of the primary carcinoma through splenec-tomy at different intervals after intrasplenic inoculation proved that the hepatic metastatic mechanism was not due to mechanical pressure but occurred spontaneously. This experimental model provides a useful means for studying the mechanism and prevention of hepatic metastasis.

A novel animal model for in vivo study of liver cancer metastasis

AIM:To establish an animal model with human hepatocyte-repopulated liver for the study of liver cancer metastasis.METHODS:Cell transplantation into mouse livers was conducted using alpha-fetoprotein(AFP)-producing hu-man gastric cancer cells(h-GCCs) and h-hepatocytes as donor cells in a transgenic mouse line expressing urokinase-type plasminogen activator(uPA) driven by the albumin enhancer/promoter crossed with a severe combined immunodeficient(SCID) mouse line(uPA/SCID mice).Host mice were divided into two groups(A and B).Group A mice were transplanted with h-GCCs alone,and group B mice were transplanted with h-GCCs and h-hepatocytes together.The replacement index(RI),which is the ratio of transplanted h-GCCs and h-hepatocytes that occupy the examined area of a histological section,was estimated by measuring h-AFP and h-albumin concentrations in sera,respectively,as well as by immunohistochemical analyses of h-AFP and human cytokeratin 18 in histological sections.RESULTS:The h-GCCs successfully engrafted,repopulated,and colonized the livers of mice in group A(RI = 22.0% ± 2.6%).These mice had moderately differentiated adenocarcinomatous lesions with disrupted glandular structures,which is a characteristics feature of gastric cancers.The serum h-AFP level reached 211.0 ± 142.2 g/mL(range,7.1-324.2 g/mL).In group B mice,the h-GCCs and h-hepatocytes independently engrafted,repopulated the host liver,and developed colonies(RI = 12.0% ± 6.8% and 66.0% ± 12.3%,respectively).h-GCC colonies also showed typical adenocarcinomatous glandular structures around the h-hepatocyte-colonies.These mice survived for the full 56 day-study and did not exhibit any metastasis of h-GCCs in the extrahepatic regions during the observational period.The mice with an h-hepatocyte-repopulated liver possessed metastasized h-GCCs and therefore could be a useful humanized liver animal model for studying liver cancer metastasis in vivo.CONCLUSION:A novel animal model of human liver cancer metastasis was established using the uPA/SCID mouse line.This model could be useful for in vivo testing of anti-cancer drugs and for studying the mechanisms of human liver cancer metastasis.

Cytotoxic Effect of Chitosan Based Nanocomposite Synthesized by Radiation： In Vitro Liver and Breast Cancer Cell Line

A silver nanoparticle （AgNP） is likely to provide an attractive object for combining a variety of biochemical properties with great therapeutic potential by using radiation. The present study explores the ICs0 value of chitosan-poly （vinyl alcohol） hydrogel （Cs/PVA） and Ag-doped chitosan-poly （vinyl alcohol） （Cs/PVA/Ag） nanocomposite in view of their anticancer application. The aim was to develop （Cs/PVA） based hydrogel synthesized by gamma radiation which could behave both as a nanoreactor for Ag nanoparticle with promising anticancer applications. The （Cs/PVA/Ag） nanocomposite was confirmed by FTIR （Fourier transform infrared） spectroscopy, XRD （X-ray diffraction） and EDX （energy dispersive X-ray） analysis. The anti-cancer activity of the prepared nanocomposites was demonstrated in human liver cancer cell line （HEPG2） and breast cancer cell lines （MCF7）. It has significant effects against human liver cancer cell line HEPG2 compared to breast cancer cell line MCF7. Further quantitative analysis on the molecular and protein levels is still required to confirm the impact of chitosan on genotoxic effect before reaching a final conclusion and starting its biomedical application.

Estabishment of A Human Liver Cancer Cell Line Transfected with IL-2 cDNA and Its Biologic Activity

Objective To obtain IL 2 gene transfected human liver cancer cells and study IL 2 expression and its biologic activity in vivo. Methods\ Human liver cancer cells SMMC 7721 were cocultured with recombinant retroviral vector LNC IL 2,and screening was performed in G418 medium.The exogenous IL 2 cDNA at the DNA,RNA,and protein levels were determined by using dot hybridization,PR PCR and MTT methods respectively.The tumorigenesis and antitumorigenesis of the screened liver cancer cell with subcutaneous injection in nude mice were observed. Results and Conclusion\ The IL 2 cDNA was successfully integrated into SMMC 7721 cell genomic DNA and continuously expressed for more than 88 days.Subcutaneous vaccination of the nude mice with transfected cells revealed an obvious suppression of its tumorigenicity,and could induce antitumor activity in vivo. \ \

Zinc protoporphyrin IX enhances chemotherapeutic response of hepatoma cells to cisplatin

AIM: To investigate the effect of zinc protoporphyrin IX on the response of hepatoma cells to cisplatin and the possible mechanism involved.

Differential Proteomics in Malignant and Normal Liver Cell Lines

Objective： To detect differential protein expression in malignant and normal liver cell lines in vitro using the SELDI ProteinChip platform, for investigating the pathogenesis of liver cancer. Methods： Two cell lines, human normal liver cell line L02 and hepatoma cell line SMMC-7721 were cultured routinely, harvested in good condition and lysed. After quantification, the supernatant of the lysate was tested by IMAC3 （Immobilized Mental Affinity Capture） and WCX2 （Weak Cation Exchange） chips on the SELDI-TOF-MS ProteinChip reader. Results： Protein expression differed between the malignant and normal liver cell lines. A total of 20 differentially expressed proteins were found, among which, 7 were captured by the IMAC3 chip and 14 by the WCX2 chip. Peaks at 5,419, 7,979 and 11,265 Da were higher and at 8,103, 8,492, 10,160 and 11,304 Da lower in SMMC-7721 cells by the IMAC3 chip; peaks at 7,517, 7,945 and 7,979 Da were higher and at 5,061, 5,551, 5,818, 7,439, 9,401,10,100, 10,312, 11,621, 11,662, 11,830 and 12,772 Da lower in SMMC-7721 cells by the WCX2 chip. Interestingly, both chips captured the 7,979 Da peak. In addition, the 11,081 Da peak corresponded precisely with the molecular mass of the calcium binding protein S100A10, which may participate in the formation of liver cancer in association with p36. Conclusion： Detecting differential protein expression in malignant and normal liver cell lines using the SELDI ProteinChip platform was simple, sensitive and repeatable. The results we obtained can serve as a basis for investigating the pathogenesis of liver cancer and aid the discovery of new therapeutic targets.

COL14A1 promotes self-renewal of human liver cancer stem cells through activation of ERK signaling

Objective:Liver cancer stem cells(CSCs)are the culprits of hepatocellular carcinoma metastasis and recurrence.Only by eliminating tumor stem cells can malignant tumors be fundamentally cured.This study aimed to identify the role and underlying mechanism of aberrant Collagen Type XIV Alpha 1 Chain(COL14A1)overexpression in liver CSCs,and improve understanding of the molecular basis of hepatocellular carcinoma metastasis and recurrence.Methods:First,quantitative real-time polymerase chain reaction was used to confirm aberrant high-expression of COL14A1 in liver CSCs.Next,interference experiments were performed to determine the key role of COL14A1.To explore the mechanism of COL14A1 overexpression in liver CSCs,putative microRNA(miRNAs)targeting COL14A1 were analyzed using the miRTarBase database.Next,quantitative real-time polymerase chain reaction,western blotting,and luciferase reporter assays were performed to verify the interaction between miR-7108-3p and COL14A1.Lastly,key target proteins of the COL14A1-extracellular-regulated signal kinase(ERK)signaling pathway were identified through western blotting analysis.This study was approved by the Ethics Committee of Shanghai Fourth People’s Hospital,Tongji University School of Medicine,China(approval No.2019tjdx17)on February 21,2019.Results:COL14A1 is abnormally highly expressed in liver CSCs,which is necessary for liver CSCs to maintain their self-renewal capability.Mechanistically,COL14A1 is post-transcriptionally regulated by miR-7108-3p in a negative manner.Low expression of miR-7108-3p increased translation of COL14A1,which subsequently activated ERK signaling,ultimately maintaining the self-renewal and stem cell-like properties of liver CSCs.Conclusion:COL14A1,which is negatively regulated by miR-7108-3p,was found to play a crucial role in maintaining the selfrenewal and stem cell-like properties of liver CSCs through activation of ERK signaling.

The therapy and mechanisms of replication-deficient recombinant adenovirus Ad-p14ARF in hepatocellular carcinoma

Objective: To study the therapeutic effect and mechanisms of recombinant adenovirus Ad-p14ARF in hepatocel- lular carcinoma cell lines. Methods: Morphology and trypan blue assay were adopted to evaluate the proliferation of different liver cancer cells after Ad-p14ARF infection. Cell apoptosis was confirmed by detecting phosphatidylserine (PS) externaliza- tion with Annexin V/PI double staining. Western blotting assay analyzed the expression of related proteins. Subcutaneous tumor model of BEL7402 was established to evaluate the therapeutic ability of Ad-p14ARF. Results: Ad-p14ARF suppressed cell growth, proliferation and promoted cell apoptosis of cancer cell lines with different genetic background. Ad-p14ARF in- hibited growth of liver cancer cells (HepG2, BEL7402) in a dose-dependent manner. Ad-p14ARF leaded to overexpression of Bax and p21, which were the downstream regulating genes of p53. Ad-p14ARF suppressed tumor growth significantly in the experimental therapy in nude mice bearing subcutaneous tumor of BEL7402. Conclusion: P14ARF gene is a powerful tumor suppressor gene to be used in cancer gene therapy. It may play an important role in gene therapy against the malignancies in the future.

Controversies in and challenges to our understanding of hepatitis C

Discovered in 1989, the hepatitis C virus (HCV) continues to cause signifi cant morbidity and mortality world-wide despite a huge research commitment to defining and understanding the virus and the disease it causes. This paper discusses a number of areas where progress in the management of the HCV have not kept pace with the scientifi c understanding of the HCV. It is suggested that in the fi elds of HCV prevention and providing access to treatment, practice falls short of what could be achieved. The role of alcohol in the pathogenesis of HCV liver injury is discussed. Discrimination against those with HCV infection and particularly those in prison settings fails to match good clinical practice. The complicated processes of sharing information between specialty groups is also discussed in an attempt to optimise knowledge dissemination in this field.

Com bined effect of alphafetoprotein antisense oligodeoxynucleotidesand 5-fluorouracil on human hepato ma cell growth

Objective To investigate the effect of alpha fetoprotein (AFP) antisense phosphorothioate oligodeoxynucleotides in combination with 5 fluorouracil (5 FU) on the growth of BEL 7404 human hepatoma cells in vitro Methods Phosphorothioate oligodeoxynucleotides were synthesized by β cyanoethyl phosphoramidite chemistry AFP gene expression in human hepatoma cells was determined by avidin biotin peroxidase complex (ABC) immunocytochemical method Cell growth in the presence or absence of experimental agents was measured using 3 (4, 5 dimethylthiazol 2 yl) 2, 5 dipheny ltetrazolium (MTT) microculture tetrazolium assay Results AFP antisense oligomers markedly suppressed the growth of BEL 7404 human hepatoma cells in vitro by sequence specific blocking of the AFP gene expression in the cells (P<0 05) 5 FU also inhibited the hepatoma cell growth in a dose dependent manner when used alone (P<0 05) The combined treatment with AFP antisense oligomers and 5 FU showed significantly enhanced hepatoma cell growth inhibition than either AFP antisense or 5 FU treated cells alone (P<0 05) Conclusion Combined use of AFP antisense oligomers and 5 FU could more effectively inhibit the growth of BEL 7404 human hepatoma cells in vitro

上期临床病案讨论结果

上期临床病案讨论结果术前诊断左上腹巨大包块：①腹膜后包块（嗜铬细胞瘤？）；②左肝肿瘤？手术情况于１９９４年５月７日在静脉复合麻醉下取双侧肋缘下人字形切口行剖腹探查术。术中见肝脏正常，未扣及结节或肿块，但在左肝下缘与胃小弯之间发现一凸出的巨大肿块。切开...