1.The effect of Tet-on gene regulated HIF-1?expression on hepatoma cell apoptosis in vitro
Zongquan XU ; Xiaoping CHEN ; Wanguang ZHANG ; Changhai LI ; Gaopeng LI
Chinese Journal of General Surgery 2000;0(11):-
Objective To investigate the effects of Tet-on gene regulated HIF-1?expression on cell apoptosis of hepatoma cells in vitro. Methods The apoptosis of hepatocellular carcinoma cell lines HePG2 was measured after HIF-1?expression was activated in HePG2 in vitro by Tet-on expression system. Results The amplified products were confirmed as the cDNA of HIF-1?by DNA sequencing, and obtained pTHE-HIF-1?was verified by endonuclease digestion. The HePG2Tet-on cells express HIF-1?constantly. After being incubated under different concentrations of doxycycline for 48 h, HIF-1?gene could inhibit HePG2 cell apoptosis. The apoptosis rates were 59. 6%、50. 9%、38. 1%、30. 5%、23. 9%、18. 3% respectively when the concentrations of doxycycline were 0?g/ml,0. 02?g/ml,0. 2?g/ml、1?g/ml、2?g/ml,5?g/ml (P
2.The effect of HBx gene on the apoptosis of hepatic cells.
Lu, YE ; Junying, QI ; Gaopeng, LI ; Deding, TAO ; Shihui, SONG
Journal of Huazhong University of Science and Technology (Medical Sciences) 2007;27(2):167-9
To study the effect of HBx gene on the apoptosis of the cell lines (L02, HepG2) and the interaction between HBx and X-linked inhibitor of apoptosis protein (XIAP), the apoptosis of pcDNA3.1-HBx transiently transfected cell lines (L02, HepG2) was detected by flow cytometry and the mRNA expression of XIAP was assayed by real-time RT-PCR. Our study showed (1) the morphology of L02/pcDNA3.1-HBx was changed and the appearance of the cells mimicked that of HepG2 cells; (2) HBx gene could be detected in L02/pcDNA3.1-HBx and HepG2/ pcDNA3.1-HBx; (3) the apoptosis rate of L02/pcDNA 3.1-HBx was higher than that of L02 cells (P<0.01) and the apoptosis rate of HepG2/pcDNA3.1-HBx was lower than that of HepG2 cells (P<0.05); (4) the XIAP expression in L02 was about 3 times that in L02/pcDNA3.1-HBx cells (P<0.01), and the expression of XIAP in HepG2/pcDNA3.1-HBx was about 4 times that in HepG2 (P<0.01). It is concluded that HBx gene may promote the apoptosis of normal hepatocytes and inhibit the apoptosis of cells of hepatic carcinoma by regulating the expression of XIAP.
3.Inhibitory activities of 3-O-β-chacotriosyl pentacyclic triterpenoids against the entry of H5 N1 influenza viruses in vitro
Gaopeng SONG ; Xintian SHEN ; Sumei LI ; Yibin LI ; Jihong FAN ; Qianqian LIANG ; Shuwen LIU
Chinese Pharmacological Bulletin 2015;(5):647-654
Aim To study the inhibitory activities of potential new anti-influenza virus agents,3-O-β-chaco-triosyl pentacyclic triterpenoids against the entry of H5N1influenza viruses.Methods Three target com-pounds were designed and synthesized structurally re-lated to the lead compound 3-O-β-chacotriosyl dioscin derivative (1 )with inhibitory activities against H5N1 influenza viruses.The inhibitory activities of these tar-get compounds were tested at a cellular level pseudo vi-rus system targeting H5N1 influenza viruse entry.Re-sults All the compounds 1 a,1 b and 1 c showed po-tent inhibitory activities against the entry of A/Thai-land/Kan353/2004 pseudo virus into the target cells, of which compound 1 b showed the best inhibitory activ-ity with an IC50 value of (1.25 ±0.22)μmol·L-1. Conclusion The SARs analysis of these compounds indicated that replacement of the aglycone moiety of compound 1 with pentacyclic triterpenoids could in-crease antiviral activity.Different types of pentacyclic triterpen as aglycone residue had the significant influ-ence on the inhibitory activity (1 b >1 c >1 a),sug-gesting ursane type of triterpenes was superior to the two other kinds of triterpenes as aglycone residue.
4.Combination of p53 and Rb in the local nano-therapy for rabbit VX2 model of hepatic metastasis ;carcinoma:the curative effect and safety evaluation
Shengli DONG ; Lu YE ; Xinbao ZHAO ; Lujing LI ; Xiaolin XU ; Bing OU ; Jingsheng PAN ; Gaopeng LI
Cancer Research and Clinic 2016;28(9):582-585,589
Objective Combination of nanoparticle with p53 and Rb gene therapy by gene targeting was applied to investigate its curative effect and safety evaluation on colorectal rabbit hepatic VX2 metastasis for tumor eradication and survival enhancement. Methods Recombinant expressing plasmids harboring wild type p53 and Rb were cotransferred or transferred separately to the rabbit hepatic VX2 metastasis by the emulsion of PLL-nHAP nanoplex and lipodiol through the hepatic artery in a tumor target manner. Subsequent co-expressions of p53 and Rb protein within the treated tumors were detected by Western blot and in situ analysis of confocal laser scanning microscope. The therapeutic effect was evaluated by the tumor growth velocity and the survival time of animals. Eventually, investigations of liver function were applied to evaluate the safety of the process. Results With safe procedure for the rabbits liver function, both p53 and Rb local nano-therapy showed favorable anti-tumor effects and increased animal survival time. p53+Rb local nano-therapy could significantly inhibit hepatic VX2 metastasis and enhance the animal survival time compared with p53 local nano-therapy or Rb local nano-therapy. Local nano-therapy showed no significant influence to animal liver function. Conclusions Rb can work synergistically with p53 in the combined therapy mediated by PLL-nHAP nanoplex to augment the anti-tumor effect. The local nano-therapy with p53 and Rb is likely to be an effective and safe anti-tumor therapy for hepatic colorectal metastasis.
5.The roles of four multi-drug resistance proteins in hepatocellular carcinoma multidrug resistance.
Gaopeng, LI ; Xiaoping, CHEN ; Qi, WANG ; Zongquan, XU ; Wanguang, ZHANG ; Lu, YE
Journal of Huazhong University of Science and Technology (Medical Sciences) 2007;27(2):173-5
The roles of multi-drug resistance protein 1 (MDR1), multi-drug resistance related protein 1 (MRP1), lung resistance protein (LRP) and breast cancer resistance protein (BCRP) in the multi-drug resistance (MDR) of hepatocellular carcinoma (HCC) were studied. By exposing HepG2 cell line to progressively increased concentrations of adriamycin (ADM), HepG2 multi-drug resistant subline (HepG2/ADM) was induced. The MDR index of HepG2/ADM was detected by using MTT. The expressions of the four MDR proteins in the three cell lines (L02, HepG2, HepG2/ADM) were investigated at mRNA and protein levels by real-time RT-PCR and Western blot respectively. Our results showed that when the ADM concentration was under 100 microg/L, HepG2 could easily be induced to be drug-resistant. The IC(50) of the HepG2/ADM to ADM was 282 times that of HepG2. The expression of MDR1 and BCRP mRNA in HepG2/ADM cells were 400 and 9 times that of HepG2 cells respectively while there was no difference in the mRNA expressions of MRP1 and LRP. There was no difference between HepG2 and L02 cells in the mRNA expressions of the four genes. At the protein level, the expressions of MDR1, BCRP and LRP but MRP1 in HepG2/ADM were significantly higher than those of HepG2 and L02. Between HepG2 and L02, there was no difference in the expressions of four genes at the protein level. HepG2/ADM is a good model for the study of MDR. The four genes are probably the normally expressed gene in liver. The expressions of MDR1 and BCRP could be up-regulated by anti-cancer agents in vitro. The MDR of HCC was mainly due to the up-regulation of MDR1 and BCRP but MRP1 and LRP. These findings suggest they may serve as targets for the reversal of MDR of HCC.
6.The role of multi-drug resistance related proteins in hepatocellular carcinoma multidrug resistance cell line HePG2/ADM
Gaopeng LI ; Xiaoping CHEN ; Qi WANG ; Zongquan XU ; Wanguang ZHANG ; Lu YE
Chinese Journal of General Surgery 2000;0(11):-
0. 05). At the protein level, the expressions of MDR1, BCRP and LRP in HePG2/ADM were significantly higher than HePG2 and L02 ( FMDR1= 28. 68, FBCRP= 18. 60, FLRP= 6. 28, P 0. 05). Conclusion The four genes are probably the normally expressed genes in the liver. The expression of MDR1 and BCRP could be up-regulated by anticancer agents in vitro. MDR1 and BCRP may play very important roles in the MDR of HePG2/ADM. However, MRP1 and LRP may have no relations with the multi-drug resistance of HePG2/ ADM cell lines.
7.The Effect of HBx Gene on the Apoptosis of Hepatic Cells
Lu YE ; Junying QI ; Gaopeng LI ; Deding TAO ; Shihui SONG
Journal of Huazhong University of Science and Technology (Medical Sciences) 2007;27(2):167-169
To study the effect of HBx gene on the apoptosis of the cell lines (L02, HepG2) and the interaction between HBx and X-linked inhibitor of apoptosis protein (XIAP), the apoptosis of pcDNA3.1-HBx transiently transfected cell lines (L02, HepG2) was detected by flow cytometry and the mRNA expression of XIAP was assayed by real-time RT-PCR. Our study showed (1) the morphology of L02/pcDNA3. 1-HBx was changed and the appearance of the cells mimicked that of HepG2 cells; (2) HBx gene could be detected in L02/pcDNA3.1-HBx and HepG2/pcDNA3.1-HBx;(3) the apoptosis rate of L02/pcDNA 3.1-HBx was higher than that of L02 cells (P<0.01) and the apoptosis rate of HepG2/pcDNA3. 1-HBx was lower than that of HepG2 cells (P<0.05); (4) the XIAP expression in L02 was about 3 times that in L02/pcDNA3.1-HBx cells (P<0.01), and the expression of XIAP in HepG2/pcDNA3. 1-HBx was about 4 times that in HepG2 (P<0.01). It is concluded that HBx gene may promote the apoptosis of normal hepatocytes and inhibit the apoptosis of cells of hepatic carcinoma by regulating the expression of XIAP.
8.3-O-β-chacotriosyl benzyl ursolate inhibits entry of H5N1 influenza virus into target cells.
Gaopeng SONG ; Xintian SHEN ; Yibin LI ; Yushan ZHENG ; Ping XIONG ; Shuwen LIU
Journal of Southern Medical University 2015;35(6):789-794
OBJECTIVETo study the inhibitory activities of 3-O-β-chacotriosyl benzyl ursolate and its derivatives as potential new anti-influenza virus agents against the entry of H5N1 influenza viruses into the target cells.
METHODSFour target compounds were designed and synthesized, which were structurally related to the lead compound 3-O-β-chacotriosyl methyl ursolate (1). The inhibitory activities of these compounds were tested at a cellular level psuedovirus system targeting H5N1 influenza viruse entry.
RESULTS AND CONCLUSIONThe compounds 1b, 1c and 1d showed potent inhibitory activities against the entry of A/Thailand/Kan353/2004 pseudovirus into the target cells, and among them compound 1d showed the strongest inhibitory activity with an IC50 value of 0.96 ± 0.10 µmol/L. The structure-activity relationship analysis of these compounds indicated that when 17-COOH of ursolic acid was esterified, introduction of Me groups rather than aryl groups more strongly enhanced the inhibitory activity. Changing 17-COOH of ursolic acid into amide could increase the antiviral activity and decrease the cytotoxicity of the compounds in MDCK cells.
Animals ; Antiviral Agents ; chemistry ; Dogs ; Influenza A Virus, H5N1 Subtype ; drug effects ; physiology ; Madin Darby Canine Kidney Cells ; Structure-Activity Relationship ; Triterpenes ; chemistry ; Virus Internalization ; drug effects
9.Effect of microneedle radiofrequency combined with supramolecular salicylic acid on facial photoaging
Lanlan JIANG ; Lan GE ; Yaoying LI ; Gaopeng LIANG ; Zhiqiang SONG
Chinese Journal of Medical Aesthetics and Cosmetology 2023;29(2):120-125
Objective:To explore the efficacy and safety of microneedle radiofrequency combined with supramolecular salicylic acid in improving facial photoaging.Methods:Thirty patients treated for facial photoaging were randomly divided into the combined treatment group and the microneedle radiofrequency group. All patients were female, aged between 30 and 50 years, with an average age of (39.4±4.1) years. The combined treatment group was treated with microneedle radiofrequency combined with supramolecular salicylic acid, and the Microneedle radiofrequency group was treated with microneedle radiofrequency alone. The patients were followed up 3 days, 7 days, 14 days, 28 days and 3 months after treatment. The results of VISIA, skin physiology and patients' satisfaction were used to compare the two groups of patients after treatment.Results:After 3 months of treatment, the skin pores (11.98±2.14 vs. 15.54±1.52), brown spots (12.40±1.85 vs. 15.84±1.42), ultraviolet spots (6.74±0.87 vs. 11.20±1.70), skin physiological status (transepidermal water loss): 11.84±1.80 vs 13.09±1.96 g/(h·m) 2, stratum corneum water content: 84.91±2.86 % vs 80.29±3.58 %, melanin index: 110.07±15.02 vs. 122.30±9.97, erythema index: 220.43±19.69 vs. 236.30±16.55), elasticity (75.98±3.94 vs. 69.89±3.58), epidermal thickness (1401.33±178.43 vs. 1217.13±139.77), skin color improvement (effective rate 86.7% vs. 40.0%) and patients' satisfaction (total satisfaction was 93.3% vs 67.0%) in the combined treatment group were significantly superior than those in the microneedle radiofrequency group, and the differences were statistically significant ( P<0.05). The reaction of moderate and severe erythema (the incidence rate 40.0% vs. 86.7%) and edema (the incidence rate 26.7% vs. 80.0%) in the combined treatment group was also significantly lower than that in the microneedle radiofrequency group ( P<0.05). No adverse pigmentation was reported in both groups after treatment. Conclusions:The combination of microneedle radiofrequency and supramolecular salicylic acid in the treatment of facial photoaging has definite clinical effect and high safety, which is worthy of clinical application.
10.Effects and Mechanisms of Combined Application of Molecular Targeted Drugs on Proliferation of Hepatocellular Carcinoma SK-Hep-1 Cells
Xiaoxia ZHU ; Yuqi JIA ; Chang LIU ; Tao GONG ; Gaopeng LI ; Hongwei ZHANG ; Baofeng YU
Cancer Research on Prevention and Treatment 2022;49(11):1126-1133
Objective To study the effects and mechanisms of molecular targeted drug combination on multi-driven proliferation hepatocellular carcinoma SK-Hep-1 cells. Methods Four molecular targeted drugs (HG6-64-1, Dasatinib, Crizotinib, and Sunitinib) were used to treat SK-Hep-1 cells, and the monophasic kinetic analysis curve and two-phase analysis curve were drawn. Western blot analysis was used to detect the effects of the above drugs on key signaling pathways in SK-Hep-1 cells. MTT assay was used to detect the effects of the above drugs and their combination on the proliferation of SK-Hep-1 cells. Results Compared with the monophasic kinetic analysis curve, the biphase analysis curve could better fit the effects of molecular targeted drugs on SK-Hep-1 cells, which predicted that the combination of HG6-64-1, Dasatinib, and MK-2206 could effectively inhibit the proliferation of SK-Hep-1 cells. Conclusion Two-phase kinetic analysis can quantitatively describe the response of multi-driven proliferation hepatocellular carcinoma SK-Hep-1 cells to molecular targeted therapy. The combination of HG6-64-1, Dasatinib, and MK-2206 is a potential drug combination for the treatment of hepatocellular carcinoma.