1.Establishment and characterization of a cell line derived from human ovarian mucinous cystadenocarcinoma
Qihui WAN ; Dagang XU ; Zongquan LI
Chinese Journal of Obstetrics and Gynecology 2001;0(07):-
Objective To establish a cell line of human ovarian cancer, and study its characterization. Methods The cell line was established by the cultivation of subsides walls, and kept by freezing. The morphology was observed by microscope and electromicroscope. The authors studied its growth and propagation, the agglutination test of phytohemagglutinin (PHA), the chromosome analysis, heterotransplanting, immuno histochemistry staining, the analysis of hormone, the pollution examination and the test of sensitivity to virus etc. Results A new human ovarian carcinoma cell line, designated ovarian mucinous cystadenocarcinoma 685 (OMC685), was established from mucinous cystadenocarcinoma. This cell line had subcultured to 91 generations, and some had been frozen for 8 years and revived, still grew well. This cell line possessed the feature of glandular epithelium cancer cell. The cells grew exuberantly, and the agglutinating test of PHA was positive. Karyotype was subtriploid with distortion. Heterotransplantations,alcian blue periobic acid schiff (AbPAS),mucicarmine,alcian blue stainings, estradiol (E 2) and progesterone were all positive. Without being polluted, it was sensitive to poliovirus Ⅰ, adenovirus 7 and measles virus. Conclusions OMC685 is a distinct human ovarian tumous cell line.
2.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
3.Effects of HIF-1α expression regulated by Tet-on gone expression system on hepatoma cell proliferation and cell cycle in vitro
Ronglong XIA ; Zongquan XU ; Ruiqin ZHANG ; Xiaoping CHEN
Chinese Journal of General Surgery 2010;25(10):834-837
Objective To investigate the effects of HIF-1α expression regulated by Tet-on gene expression system on cell proliferation and cell cycle of hepatoma cells in vitro. Methods The change of human hepatocellular carcinoma cell lines HepG2 cell cycle and cell proliferation was measured after HIF-1 α expression of HepG2 in vitro was regulated by Tet-on expression system. Results Amplified products were confirmed as the cDNA of HIF-1α by DNA sequencing, and pTRE-HIF-1α obtained by edonuclease digestion,capable of expression in HepG2 Tet-on cells. After being incubated under different concentrations of doxycycline for 48 h, MTT assays showed that up-regulation of HIF-1α expression increased HepG2 cell proliferation activities. The cell index of S and G2/M phase was significantly higher and that of G0/G1 phase reduced with the increasing concentrations of doxycycline. The mRNA expression of Cyclin A increased with the increasing concentrations of doxycycline ( P < 0. 001 ), CyclinD1 and CyclinE did not change ( P >0. 05). Conclusion HIF-1 α gene promotes cell proliferation and cell cycle of hcpatoma cells in vitro, and this effects increased with the increasing of HIF-1α expression possibly through influencing the expression of CyclinA.
4.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.
5.STUDIES ON THE DIAGNOSIS OF HUMAN FILARIASIS BY INDIRECT FLOURESCENT ANTIBODY TEST(IFAT) AND IMMUNOENZYMATIC STAINING TECHNIQUE(IEST) WITH HOMOLOGOUS AND HETEROLOGOUS ADULT FILARIAL ANTIGENS
Yunhe LI ; Yonde HU ; Wenmei XU ; Zongquan WU ; Guangbao YANG ; Shuilong YIN ; Wei LUO ; Wei GONG ;
Chinese Journal of Parasitology and Parasitic Diseases 1987;0(01):-
This article deals with the diagnostic values of IFAT and IEST in human filariasis with both frozen sections of Brugia malayi and Setaria cervi adult worms as antigens. The average positive and the false positive rates of IFAT with two antigens were 84.9-97.1%(45/53-102/105)and 2.9-9.7%(1/35-3/31) respectively, while no cross-reaction was observed in subjects infected with ascaris or hookworm. The average positive and the false positive rates of IEST with two antigens were 94.3-94.9% (99/105-56/59) and 0-2.9% (0/35-l/35) respectively. Both IFAT and IEST with two antigens for the diagnosis of human filariasis were considered to be of higher sensitivity and specificity, frozen sections of adult Setaria cervi being more economical and effective.
6.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.
7.COMPARISON OF IMMUNOENZYMATIC STAINING TECHNIQUE (IEST), DOUBLE GLUING STRIP CIRCUMOVAL PRECIPITIN TEST(DGS-COPT) AND CONVENTIONAL CIRCUMOVAL PRECIPITIN TEST(CV-COPT) FOR DIAGNOSIS OF SCHISTOSOMIASIS JAPONICA
Yunhe LI ; Yonde HU ; Wenmei XU ; Guangbao YANG ; Shuilong YIN ; Wei GONG ; Jiayong ZHU ; Zongquan WU ; Wei LUO
Chinese Journal of Parasitology and Parasitic Diseases 1987;0(04):-
IEST, DGS-COPT and CV-COPT using lyophilized ova of schistosoma japonicum were performed on sera from 120 cases of schistosomiasis japonica, 120 cases of schis-tosomiasis japonica 3-8 years after being cured with praziquantel and 120 healthy individuals by single-blind method. The sensitivity and specificity of IEST was 91.7% and 95.8% respectively which were significantly higher than that of both DGS-COPT and CV-COPT. The negative conversion rate of cured patients was 70.8% with IEST, 80.8% with DGS-COPT and 81.7% with CV-COPT. The results showed that IEST has higher diagnostic value for schistosomiasis than both COPT. DGS-COPT has the same diagnostic value as CV-COPT, however, it was easy to perform and time-saving, thus it might be applied in the fields for practical purposes.
8.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):173-175
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 μg/L, HepG2 could easily be induced to be drug-resistant. The IC50 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 liven 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.