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1.
Chinese Pharmacological Bulletin ; (12): 366-372, 2016.
Article in Chinese | WPRIM | ID: wpr-487210

ABSTRACT

Aim To investigate the effect of Rhein on the movement and invasiveness of human ovarian carci-noma cells with directional high lymphatic metastasis SKOV3-PM4 cells and explore the role of Rac1/LIMK1/cofilin signaling pathway. Methods Migration assay and invasion assay were used to observe the effect of Rhein on the metastatic and invasive ability of SK-OV3-PM4 cells in vitro. The effect of Rhein on the morphology and cytoskeleton ultrastructure of ovarian cancer cells was observed by laser scanning confocal microscope and scanning electron microscope. The protein expression level of Rac1,LIMK1,PAK1 and co-filin were detected by Western blot, respectively. Re-sults Rhein inhibited the abilities of cell invasion and migration of SKOV3-PM4 cells,and the inhibitory rate increased along with the increase of the concentration and treatment duration. After treated with 8. 80 μmol· L-1 ,17. 60 μmol · L-1 , 26. 40 μmol · L-1 of Rhein for 24 h,the abilities of migration and invasion of SK-OV3-PM4 cells were inhibited ( P <0. 05 ); the mor-phology and cytoskeleton ultrastructure of SKOV3-PM4 cells were changed, cellular pseudopod reduced, cell microfilament fractured and its distribution disordered, plasma membrane was uneven and cell gap widened . After treatment of Rhein and Rac1 inhibitor , Rac1 protein expression and the expression of P-LIMK1 , P-PAK1 and P-cofilin notably decreased in a dose-de-pendent manner compared with the control group ( P<0. 05 ) . After Rhein and Rhein plus Rac1 inhibitor treatment ,P-LIMK1, P-cofilin, P-PAK1 protein levels of SKOV3-PM4 cells significantly decreased compared with the control group , and the group of Rac1 inhibi-tor plus Rhein treatment, the phosphorylated protein decreased more significantly ( P <0. 05 ) . After Rac1 activator plus Rhein treatment, phosphorylated protein expression of P-LIMK1 ,P-PAK1 and P-cofilin upregu-lated significantly ( P <0. 05 ) . Conclusions Rhein may be a potential inhibitor of Rac1 and can inhibit the migrating and invasive capabilities of directional high lymphatic metastasis SKOV3-PM4 cells through down-regulating the phosphorylation of Rac1/LIMK1 /cofilin pathway associated protein.

2.
Chinese Pharmacological Bulletin ; (12): 1175-1180, 2016.
Article in Chinese | WPRIM | ID: wpr-495772

ABSTRACT

Aim To synthesize 8-bromo-ethoxy Rhein and investigate its mechanisms and inhibition effect on hepatitis B surface antigen ( HBsAg ) and e antigen ( HBeAg) in HepG2.2.15 cells.Methods 8-bromo-ethoxy Rhein was synthesized based on the chemical structure of Rhein , and its structure was identified by IR,1 H-NMR and 13 C-NMR spectra.MTT assay was used to test the inhibitory effect of 8-bromo-ethoxy Rhein on HepG2.2.15 cells.After the cells treatment by 8-bromo-ethoxy Rhein , the HBsAg and HBeAg in cell supernatant were detected by ELISA .The expres-sion of hepatitis B virus X gene ( HBx) was detected by Western blot .The cell cycles were examined with flow cytometry.The intracellular free calcium concentration was detected by laser scanning confocal microscopy . Results The structure of 8-bromo-ethoxy Rhein was confirmed by IR,1 H-NMR and 13 C-NMR.MTT results showed that synthetic product and Rhein could inhibit the cell proliferation in HepG2.2.15 cells.After trea-ted with 8-bromo-ethoxy Rhein and Rhein for 72 h,the half inhibitory concentration 50%( IC50 ) was 14.29 mg? L-1 and 11.59 mg? L-1 , respectively .Using non-cytotoxic dose of 8-bromo-ethoxy Rhein , the inhibitory effect on HBsAg and HBeAg was gradually enhanced with increasing 8-bromo-ethoxy Rhein concentration . The inhibitory effect of synthetic product on hepatitis B virus was better than that of Rhein .8-bromo-ethoxy Rhein could down-regulate the expression of HBx , in-tracellular calcium ion concentration and block the hepatitis B virus ( HBV ) replication.Flow cytometry results showed 8-bromo-ethoxy Rhein didn′t affect the cell cycle .Conclusions Compare with Rhein , the synthesis of 8-bromo-ethoxy Rhein shows stronger inhi-bition on hepatitis B virus in HepG2.2.15, and its mechanisms may involve down-regulating the expres-sion of HBx and reducing calcium ion concentration .

3.
Chinese Journal of Obstetrics and Gynecology ; (12): 510-516, 2014.
Article in Chinese | WPRIM | ID: wpr-453815

ABSTRACT

Objective To establish the condition cultrue cell system and co-culture cell system with SKOV3/PM4,HUVEC and to study the changes of their biological characteristics. Methods The cells of SKOV3/PM4 and HUVEC were labeled with green and red fluorescent respectively. The cell supernatant of SKOV3/PM4 and HUVEC were collected respectively as the condition medium(e.g:the cell supernatant of HUVEC cells was used as SKOV3/PM4 condition medium)and to establish the condition cultrue cell system and the co-culture cell system of the two cell lines. In the condition cultrue cell system, The morphological changes of cells were observed by HE staining to calculate the mitotic index. The ultrastructural changes of the two cells were observed by transmission electron microscopy(TEM). The growth curve of the cells was determined by methyl thiazolyl tetrazolium (MTT) assay and flow cytometry was used to analyzed the cell cycles.In the co-culture cell system, the interaction of the two cells were detected by laser scanning confocal microscope(LSCM). The expression of matrix metalloproteinase-2(MMP-2) and matrix metalloproteinase-9 (MMP-9) were detected by gelatin zymography. Results Compared with the single culture SKOV3/PM4, the cells which cultured in HUVEC condition medium showed the increase of pseudopodia and nuclear division,the mitotic index respectively were [(4.8 ± 0.8)%,(11.2 ± 0.3)%;P<0.05]. The growth rate was significantly increased. In cell cycles, it showed the declined cell ratio of G0/G1 phase, respectively[(69.4±3.6)%, (48.4±4.6)%;P<0.05] and the raised cell ratio of G2/M phase, respectively [(5.2±1.6)%, (24.9±2.2)%;P<0.05]. Compared with the single culture HUVEC,the cells which cultured in SKOV3/PM4 condition medium showed the significant morphological change and vacuolization in the cytoplasm, Nuclear division was increased and the mitotic index respectively were [(2.7±0.5)%, (5.7±0.6)%;P<0.05]. The growth rate was slightly declined. In cell cycles, it showed the raised cell ratio in G0/G1 phase, respectively [(51.4 ± 2.2)%,(79.0 ± 4.1)%;P<0.05] and the declined cell ratio in G2/M phase, respectively [(19.1±1.2)%, (3.3±0.5)%;P<0.05]. After co-culture for 48 hours, spontaneous fusion between SKOV3/PM4 and HUVEC cell was observed by the laser confocal microscope. Gelatin zymography assay showed that MMP-2 was not expressed in HUVEC cells, low-expressed in SKOV3/PM4 cells and high-expressed in the co-culture SKOV3/PM4+HUVEC cells. The expression of MMP-2 in co-culture SKOV3/PM4+HUVEC cells and SKOV3/PM4 cells respectively were 1 885 ± 84 and 1 209 ± 114 (P<0.05). But there were no MMP-9 expression in HUVEC cells, SKOV3/PM4 cells, and the co-culture SKOV3/PM4+HUVEC. Conclusion The characteristics of SKOV3/PM4 and HUVEC show significant changes after condition culture and co-culture, it may involve in the microenvironment of the cells and the intercellular crosstalk pathway.

4.
Chinese Journal of Clinical Oncology ; (24): 1155-1159, 2013.
Article in Chinese | WPRIM | ID: wpr-438257

ABSTRACT

Objective:This study aimed to investigate the effect of human lymphatic endothelial cells (HLECs) on proteins secreted by epithelial ovarian cancer (EOC) cells SKOV3-pm4 with highly directional lymphatic metastasis. Methods:The supernatants of the four groups of cultured cells (A, SKOV3;B, SKOV3+HLEC;C, SKOV3-PM4;and D, SKOV3-PM4+HLEC) were collected. The proteins of these cells were detected by antibody arrays and iTRAQ-2D-LC-MALDI-TOF/TOF/MS. The screened significantly differential proteins were further analyzed by bioinformatics and validated in the human serum and cell culture medium by ELISA. Results:Progranulin (GRN) and vascular endothelial growth factor A (VEGF-A) were upregulated between groups C and A. In addition, insulin-like growth factor binding protein-7 (IGFBP-7) and secreted protein acid rich in cysteine (SPARC) were downregulated between groups D and C. Comprehensive bioinformatics analysis revealed that IGFBP7 interacted with VEGFA. VEGF exhibited the highest expression in ovarian cancer and IGFBP7 exhibited the lowest expression compared with the serum of the normal control group. Statistically significant differences were observed between the two substances. Conclusion:The HLEC microenvironment is closely associated with directional metastasis in lymph nodes with differential proteins, including matricellular proteins and adhesion factors. In particular, the upregulation of VEGFA and GRN and the downregulation of SPARC and IGFBP7 were closely associated with the directional metastasis of EOC cells in lymph nodes.

5.
Chinese Journal of Obstetrics and Gynecology ; (12): 34-40, 2013.
Article in Chinese | WPRIM | ID: wpr-432924

ABSTRACT

Objective To investigate the efficiency of biology function of ITIH4 gene silenced by small interfering RNA (siRNA) on ovarian cancer.Methods The four pairs ITIH4 gene siRNA interference fragments(ITIH4-546,ITIH4-795,ITIH4-917 and ITIH4-1568) were designed respectively,and transfected into HO8910pm cells with ITIH4 mRNA high expression by liposomal method transiently.Quantitative PCR method was used to detect the ITIH4 mRNA expression in HO8910pm cells transfected with interference fragment.The ITIH4 917 was selected as the best silencing effect of siRNA interference fragment and then the recombinant plasmid expression vector pGPU6/GFP/Neo-shRNA-ITIH4-917 was constructed and transfected into HO8910pm cells.The stably transfected cells-pGPU6/GFP/Neo-shRNA-ITIH4-917-HO8910pm cells was obtained by screening of aminoglycoside antibiotics (G418).The experiment was divided into three groups,namely ITIH4-917 transfection group,the HO8910pm cell group transfected with pGPU6/GFP/Neo-shRNA plasmid (empty vector group),and the HO8910pm cell group transfected with pGPU6/GFP/Neo-shRNA-ITIH4-NC the plasmid (negative control group).Fluorescence quantitative reverse transcription(RT)PCR and western blot were used to detect the ITIH4 mRNA and protein expression.The cell proliferation,the cell cycle,colony formation of cells,cells migration and invasion in vitro were determined by using methyl thiazolyl tetrazolium (MTT),flow cytometry,colony formation assay and transmembrane (transwell) small chamber method [value represented by absorbance (A)],respectively.Results The fluorescent quantitative PCR results showed that the ITIH4 mRNA expression levels in ITIH4-917 HO8910pm cells was significantly lower than that in the control cells,the relative copy number was only 0.26 ± 0.15.Also the relative copy number of ITIH4 mRNA in ITIH4-917 transfection group cells was 0.34 ±0.10,it significantly lower than that in empty vector group (1.87 ±0.12,P =0.008) and negative control group (1.58 ±0.21,P =0.032) ; Western blot results showed that the ITIH4 relative expression levels of the protein in ITIH4-917 HO8910pm group cells,empty vector group and negative control group were 0.51,1.64 and 1.74,respectively,there were statistically significant differences (0.51 vs.1.64,P =0.012; 0.51 vs.1.74,P =0.014).MTT colorimetric assay showed that the proliferation of ITIH4-917 HO8910pm group cells was significantly faster than that in the empty vector group and negative control group,and there were statistically significant differences among them (P =0.001).The S ± G2/M phase cell ratio in ITIH4-917 HO8910pm group cells was 54.2%,which was significantly higher than that in the empty vector group or negative control group (26.3% and 31.3%,respectively,all P < 0.05).The colony formation rate (55.7 ± O.7) % in ITIH4-917 HO8910pm group cells was also significantly higher than that in empty vector group (29.7 ±0.9) % (P =0.037) and negative control group (31.4 ± 0.3) % (P =0.043).Migration and invasion experiments showed that cell migration in ITIH4-917 HO8910pm group cells was 0.40 ± 0.18,whicht was significantly higher than that in the negative control group or empty vector (0.30 ±0.03,P =0.031 ;0.25 ±0.03,P =0.028,respectively).Although the invasive ability of ITIH4-917 HO8910pm group cells (1.31 ±0.34) was higher than that in the control cells (1.05 ±0.68) and empty vector group (1.14 ±0.08),while there were not significant difference (P > 0.05).Conclusion It would be to promote the cell doubling time and increase the migration capability in HO8910pm cells that ITIH4 expression was down-regulating by ITIH4 mRNA interference.

6.
Chinese Journal of Radiological Medicine and Protection ; (12): 20-24, 2012.
Article in Chinese | WPRIM | ID: wpr-424841

ABSTRACT

Objective To discover radioresistance associated molecular biomarkers and its mechanism in nasopharyngeal carcinoma by protein-protein interaction network analysis.Methods Whole genome expression microarray was applied to screen out differentially expressed genes in two cell lines CNE- 2R and CNE-2 with different radiosensitivity.Four differentially expressed genes were randomly selected for further verification by the semi-quantitative RT-PCR analysis with self-designed primers. The common differentially expressed genes from two experiments were analyzed with the SNOW online database in order to find out the central node related to the biomarkers of nasopharyngeal carcinoma radioresistance. The expression of STAT1 in CNE-2R and CNE-2 cells was measured by Western blot.Results Compared with CNE-2 cells,374 genes in CNE-2R cells were differentially expressed while 197 genes showed significant differences.Four randomly selected differentially expressed genes were verified by RT-PCR and had same change trend in consistent with the results of chip assay. Analysis with the SNOW database demonstrated that those 197 genes could form a complicated interaction network where STAT1 and JUN might be two key nodes.Indeed,the STAT1-α expression in CNE-2R was higher than that in CNE-2 (t =4.96,P < 0.05).Conclusions The key nodes of STAT1 and JUN may be the molecular biomarkers leading to radioresistance in nasopharyngeal carcinoma,and STAT1-α might have close relationship with radioresistance.

7.
Chinese Journal of Obstetrics and Gynecology ; (12): 41-44, 2010.
Article in Chinese | WPRIM | ID: wpr-391500

ABSTRACT

Objective To detect the genetic transcription and protein expression level of endoplasmic reticulum aminopeptidase 1 (ERAP1) in human ovarian cancer cells and tissue and study their relationship with directional lymphatic metastasis. Methods Real-time fluorescent quantitative PCR and western blot were used to determine the expressions of ERAP1 gene and protein in the human ovarian cancer cell lines between non-directional (SKOV3) and directional highly lymphatic metastasis (SKOV3-pm2, SKOV3-pm3, SKOV3-pm4 ). Immunohistochemistry method was used to further validate the ERAP1 expressions of the transplanted ovarian tumor primarily focus and the lesions of lymph node metastasis from nude mice and the human ovarian cancer primarily focus and the lesions of lymph node metastasis. Results The expression level of ERAP1 gene and protein were down-regulated in SKOV3, SKOV3-pm2, SKOV3-pro3, SKOV3-pm4 cell sublines (0.118±0.012, 0.031±0.003,0.028±0.003, 0.016±0.005 ; 0.91± 0.33, 0.09±0.03, 0.10±0.04, 0.05+0.04; respectively), and the level of ERAP1 in SKOV3 cell was higher than those in the other three kinds of cell lines (P<0.05 ). The results showed that there were significant declining trend of expression of ERAP1 in the human ovarian cancer cell lines between non-directional and directional highly lymphatic metastasis; the transplanted ovarian tumor primarily focus and the metastasis lesions of lymph node from nude mice (143±22 vs. 97±12, P<0.05), the primarily focus (184±14) and the lesions of lymph node metastasis from human ovarian tumors (P<0.05). Conclusion The absence or down-regulated expression of ERAP1 is closely related to the metastasis and invasion of lymph node in ovarian carcinoma.

8.
Chinese Journal of Obstetrics and Gynecology ; (12): 598-602, 2010.
Article in Chinese | WPRIM | ID: wpr-387715

ABSTRACT

Objective To clone cathepsin L (CTSL) gene and construct the eukaryotic expression plasmid pcDNA3. 1-CTSL and study the relationship between CTSL and invasion and metastasis in ovarian cancer cells in vitro. Methods The total RNA was extracted from the ovarian cancer tissue and the intact cDNA of CTSL was applied by reverse transcription (RT)-PCR. The product of RT-PCR was cloned to pMD18-T vector, and subcloned to pcDNA3. 1 vector. It was tested by the enzymation and DNA sequencing.The eukaryotic expression plasmid of CTSL was introduced into HO8910 cells by liposome transfection reagent. RT-PCR was used to confirm the recombinant plasmid DNA integrated with the genomic DNA of HO8910 cells. Western blot was used to confirm the CTSL protein expression in positive clones cells. The cell growth curves, clonogenicity efficiency were observed. The cell cycles were measured by flow cytometer.The ability of invasion, metastasis and adhesion of ovarian cancer cells were detected by the matrigel invasion assay, transwell migration assay and adhesion assay, respectively. Results The results from restrictive enzyme analysis and sequencing showed that the CTSL gene was successfully inserted into pcDNA3. 1.Result from RT-PCR and western blot showed that the ovarian cancer cells which transfected by recombinant plasmid could express CTSL gene and protein. There was no difference between HO8910-CTSL and HO8910-pcDNA3. 1 cells in proliferation and adhesion ability (0.16±0.04 versus 0. 19±0. 04) of the cells (P>0.05). There was difference between HO8910-CTSL and HO8910-peDNA3.1 cells in matrigel invasion ability (0.34±0.18 versus 0.17±0.04) and metastasis ability (1.252±0.114 versus 0.486±0.027) of cancer(all P<0.05). Conclusion CTSL maybe increase the ability of invasion and metastasis of ovarian cancer cells in vitro, which may be a molecular target of blocking invasion and metastasis of ovarian cancer.

9.
Chinese Journal of Obstetrics and Gynecology ; (12): 686-690, 2009.
Article in Chinese | WPRIM | ID: wpr-392525

ABSTRACT

Objective To explore whether or not multi-drug resistance could be reversed by RNA interference the expression of Topo Ⅱα gene in epithelial ovarian cancer cell lines in vitro. Methods (1) The best silent small interference RNA (siRNA) of Topo Ⅱα gene was designed and chose and cloned into psilencer4, 1-CMV-neo vector. The psilencer4. 1-CMV-neo-Topo Ⅱα was transfected into SKOV3/DDP cell, then Topo Ⅱα siRNA (+) SKOV3/DDP cells was incubated. (2) The Tope Ⅱα mRNA and protein expression of the stability-transfecting cell lines were detected by RT-PCR and western blot method, respectively. The resistance index, the cell cycle and the cellular content of cisplatin were detected by methyl thiazolyl tetrazolium assay, the flow cytometry and high performance liquid chromatography method before and after Topo Ⅱα RNA interference in cells. Results (1) The Topo Ⅱα gene expression level in SKOV3/DDP cells could be inhibited after the plasmid DNA psilencer4, 1-CMV-neo-Topo Ⅱα transfeced. The expression level of Tope Ⅱα mRNA in Topo Ⅱα siRNA(+)SKOV3/DDP and SKOV3/DDP cells were 0 and 0.92±0.08; the expression level of Topo Ⅱα protein in Topo Ⅱα siRNA (+) SKOV3/DDP and SKOV3/DDP cells were 0.51±0. 04 and 1.95±0.09 (P<0.01). (2) The multi-drug resistance index of Topo Ⅱα siRNA (+) SKOV3/DDP cell was significantly lower compared with that in SKOV3/DDP cell (3.46 vs 5.05, P<0.05). (3) The percentage of G_0/G_1 and G_2/M phase cell in Topo Ⅱα siRNA(+) SKOV3/DDP cells were higher than that in SKOV3/DDP cells (P<0.05). (4) The content of cisplatin in Topo Ⅱα siRNA(+)SKOV3/DDP cells treated with cisplatin for 24 hours was significantly higher than that in SKOV3/DDP cell (157.20 vs 63.99 ng, P<0.05). Conclusion The results showed that the tolerance of cisplatin would be reversed by blocking the Topo Ⅱα gene expression in cisplatin-resistant epithelial ovarian cancer cells.

10.
Chinese Journal of Obstetrics and Gynecology ; (12): 854-858, 2008.
Article in Chinese | WPRIM | ID: wpr-397722

ABSTRACT

Objective To assess the suppression effect on WWOX gene in SKOV3/SB cell line by small interference RNA (siRNA). Methods Transfection of siRNA using lipofeetamine 2000 was conducted to silence WWOX gene expression, the expression levels of WWOX mRNA and protein were evaluated,and the effects on the cell cycles at 48 hours of transfection were assessed by RT-PCR, western blot and flow eytometry (FCM) respectively. The cisplatin resistance index was assayed after transfection of SKOV3/SB with siRNA by methyl thiazolyl tetrazolium(MTT) and the cisplatin concentration of SKOV3/SB cells transfected with siRNA of WWOX was measured by high performance liquid chromatography. Results(1) In SKOV3/SB cells transfected with WWOX interference fragment, whether at the mRNA or protein level, the expression of both of WWOX decreased. There was a significant difference (P<0.05) compared with SKOV3 cells and non-transfected cells. (2) After transfecfion of the WWOX interference fragment, the index of platinum resistance of SKOV3/SB decreased from 5.04 to 3.89. (3) The number of cell transfected with the WWOX interference fragment in G1 phase was increased, while that in S-phase was decreased. (4) The cisplatin concenla'ation of SKOV3/SB cells transfected with the WWOX interference fragment was increased from 9.43 ng/L to 23.45 ng/L compared with SKOV3/SB cells non-transfected with a significant difference (P<0.05). Conclusion WWOX gene may be involved in cisplatin resistance phenomenon in epithelial ovarian cancer.

11.
Chinese Pharmacological Bulletin ; (12)2003.
Article in Chinese | WPRIM | ID: wpr-554162

ABSTRACT

AIM To investigate the effects of Tea Po lyp henols on induction of apoptosis in human liver cancer cell etc. Telomerase acti vity of BEL-7404 cells was dramatically declined during the cell incuba tion with Tea Polyphenols. METHODS MTT colorimetric assay and trypan b lue exclusion method were used to examine the growth inhibition ofTea Polyphenols on BEL-7404 cells. Telomerase activity in cell was determined by PCR-ELISA method. Cell morphology and DNA gel electrophoresis were used to observe the apoptosis. RESULTS The solution of Tea Polyphenols showed dose-dependent and time-dependent inh ibitory effects on the proliferation on BEL-7404 cell line. After treatment wit h 0 2 and 0 1 g?L -1 tea polyphenols for 48 hours, cells displayed DNA ladder bands and typical morphological change of apoptosis included: ce ll shrinkage, membrane blebbing, chromatin condensation, nuclear fragmentation etc. Telomerase activit y of BEL-7404 cells was dramatically declined during the cell incubation with Tea Polyphenols. CONCLUSION Tea Polyphenols has growth inhibiti ng effect and may induce apoptosis in BEL-7404 in a certain dose range. The ant icancer mechanism of Tea Polyphenols might be related to the inhibition of telom erase activity.

12.
China Oncology ; (12)1998.
Article in Chinese | WPRIM | ID: wpr-541687

ABSTRACT

Purpose:To explore the role of terminal restric tion fragments (TRFs), telomerase activity and expression of human telomerase re verse transcriptase (hTERT) in colorectal carcinoma. Methods:The telomere length, telomerase activity and expression of hTERT were studied with Southern-blot, TRAP and immunohistochemistry, respe ctively. Results:TRFs in cancer tissue was much shorter than in the adja cent tissues and normal mucosa, and TRFs was decreased gradually along with the development in cancer stage.The expression of Telomerase in colorectal carcinoma tissue was significantly higher than that in other tissues (P

13.
Chinese Pharmacological Bulletin ; (12)1987.
Article in Chinese | WPRIM | ID: wpr-565256

ABSTRACT

Aim To investigate the effects of emodin isolated from Guangxi P.multiflorum Thunb on the expression of KU70/KU80 in hypoxic nasopharyngeal cancer CNE-1 cells and reveal the relationship between radiosensitization of emodin monomer and DNA repair genes.Methods The expression of hypoxia inducible factor-1?(HIF-1?)and DNA double-strand break repair genes(KU70/KU80)between the experimental groups and the control group under hypoxic condition was detected by the real-time fluorescence quantitative RT-PCR.Results Expression of HIF-1? was significantly increased under hypoxia condition.HIF-1? had no change after treatment with emodin alone.The expression level of KU70/KU80 was induced by radiation.Compared with radiation alone group,radiation combined hypoxia group obviously enhanced the expression of KU70/KU80.KU70/KU80 mRNA expression significantly reduced after radiation combined with emodin under hypoxic condition.Conclusion In the hypoxic environment,emodin combined with radiotherapy can effectively inhibit the expression of HIF-1 ? and DNA double-strand break repair genes(KU70/KU80),which may be its mechanism of radiosensitization.

14.
Chinese Pharmacological Bulletin ; (12)1986.
Article in Chinese | WPRIM | ID: wpr-678003

ABSTRACT

AIM To observe the effects of Oxymatrine on induction of apoptosis in human ovarian cancer SKOV3 cells. METHODS Apoptosis induced by Oxymatrine in human ovarian cancer SKOV3 cells was tested by MTT assay, fluorescent microscope, and DNA gel eletrophoresis. RESULTS SKOV3 cell viability dropped down depending on the Oxymatrine concentration and treatment time. When incubated with Oxymatrine (0 189 mmol?L -1 and 0 378 mmol?L -1 ) for 48 h, SKOV3 cells showed morphological changes associated with the characters of apoptosis under fluorescent microscope. Typical DNA ladder was found during gel eletrophoresis. CONCLUSION Oxymatrine can induce apoptosis in SKOV3 cell lines.

15.
Chinese Pharmacological Bulletin ; (12)1986.
Article in Chinese | WPRIM | ID: wpr-561337

ABSTRACT

Aim Telomerase is highly expression in most tumor cells, and it is an ideal target for cancer molecular targeting therapy. It has been proved that wogonin effectively inhibits telomerase activity and tumor cell growth in vitro. The study was to explore the inhibitory effect of wogonin on the growth of tumor and telomerase activity of implanted human ovarian cancer cell line SKOV3 in nude mice. Methods Nude mice with implanted human ovarian cancer cells SKOV3 were randomly divided into five groups, viz. the high dose group of Wogonin(600 mg?kg-1),low dose group of Wogonin(300 mg?kg-1),normal control group, cisplatin therapy group(3 mg?kg-1), and combined therapy group(cisplatin plus wogonin).The weight of nude mice and the volume of tumor were regularly measured. DNA、RNA and protein were extracted from the tumor tissue. The length of telomere was examined by Southern blot. The expression of telomerase hTERT gene was detected by RT-PCR. The telomerase activity was examined by TRAP-PCR-silver staining. Results The wogonin significantly inhibit the growth of tumor when compared with controlled group.The inhibitory rate of high dose group and low dose group were 56.67% (P=0.002) and 38.10%(P=0.019), respectively. The inhibition rate of cisplatin therapy group was 50.83%(P=0.004). The suppress rate of combined group reached 66.9% and higher than any single therapy(P=0.002). The length of telomere in different concentration groups of wogonin was the same as that in the control group.Wogonin inhibited the expression of telomerase gene hTERT and telomerase activity. The inhibition is related to the dose of wogonin. Conclusion Wogonin suppresses the growth and telomerase activity of tumor. The inhibitory effect is related to the dose of wogonin. Combination of wogonin and cisplatin increase the inhibitory rate in nude mice tumor.

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