2.Reversal of multidrug resistance of K562/ADM cells in vitro by the monomer of traditional Chinese medicine extract and investigation of its molecular mechanism
Gaojuan QIAO ; Guihai LI ; Weihua YANG ; Peie WEN ; Xia REN ; Hua FAN ; Tianhua TANG ; Haiquan REN ; Guosheng JIANG
Journal of International Oncology 2009;36(6):474-476
Objective To investigate the reversal effect of the monomer of traditional Chinese medicine on muhidrug resistance(MDR) and its possible mechanism in K562/ADM cell line in vitro. Methods With different concentrations of baicalin, geniposide administered to K562/ADM cells, the proliferation of K562/ ADM cells was detected by the MTY assay. Expression of mdr-1 mRNA, Topo Ⅱ mRNA was measured by semi-quantitive RT-PCR. Results Thatbaicalin and geniposide could increase the sensitivity of K562/ADM cells to adriamycin, multiples of reversion were 1.95 times and 1.46 times. The proliferation of K562/ADM cells was in-hibited obviously by baicalin and geniposide, the level of mdr-1 mRNA expression was down-regulated and the Topo Ⅱ mRNA was up-regulated(P<0.01 ). Conclusion Baicalin and geniposide may reverse the multi-drag-resistance of K562/ADM cells, which was related to the down-regulation of mdr-1 expression and up-reg-ulation of Topo Ⅱ beta expression.
3.Mechanism of VEGF expression and secretion in HL-60 cells induced by all-trans refinoic acid
Hua FAN ; Peie WEN ; Weihua YANG ; Xia REN ; Haitao ZHAO ; Gaojuan QIAO ; Tianhua TANG ; Haiquan REN ; Guosheng JIANG
Journal of International Oncology 2009;36(4):316-319
Objective To explore molecular mechanism of expression of vascular endothelial growth factor (VEGF) mRNA and secretion of VEGF protein in HL-60 cells induced by all-trans refinoic acid (ATRA). Methods MTr method was used to detect the proliferation of HL-60 cells induced by ATRA,cell cycle and CD11b expression in HL-60 cells were detected by flow cytometry. Expression of VEGF, c-myc, by-poxia-inducible factor(HIF)-lα, matrix metalloproteinase (MMP)-9 and MMP-2 mRNA were detected by semi-quantitative RT-PCR. VEGF protein in HL-60 cells culture supernatant was measured by ELISA before and after being induced by ATRA. Results After treatment with ATRA,the proliferation of HL-60 cells was obviously inhibited, CD11b expression increased, trend of granulocyte directional differentiation emerged, and differentiation degree was increasd(P <0. 05) ;expression level of c-my and VEGF mRNA was down-regulated (P < 0. 05), but expression level of HIF-1α mRNA was up-regulated (P < 0. 05). VEGF protein level in HL-60 cells culture supernatant was decreased by blocking the expression of MMP-9 or MMP-2(P <0. 05).Conclusion VEGF expression has positive correlation with c-myc expression,but has negative correlation with HIF-1α expression. MMP-9 and MMP-2 may be the main factors regulating VEGF secretion in HL-60 cells.
4.Molecular mechanism of different sensitivities of PML-RARα to apoptosis induced by apoptosis inducing agents
Haitao ZHAO ; Peie WEN ; Xia REN ; Weihua YANG ; Hua FAN ; Gaojuan QIAO ; Tianhua TANG ; Haiquan REN ; Kehong BI ; Guosheng JIANG
Journal of International Oncology 2009;36(5):394-397
Objective To study the molecular mechanism of different sensitivities to apoptosis induced by low concentration of As2O3 in PML-RARα negative HL-60 cells and PML-RARα positive NB4 cells. Meth-ods NB4 and HL-60 cells were cultured with As2O3 for 1 to 4 days; cell proliferation were detected by MTT method; the apoptosis was detected by flow cytometry,Bcl-2,Bax and Fas mRNA were determined by RT-PCR. Results The proliferation of NB4 cells was inhibited obviously by As2O3(1.0 μmol/L)with the induction of apoptosis( P <0.05) ,which was accompanied by the down-regulation of Bcl-2 mRNA expression( P <0.05)and the ratio of Bcl-2/Bax(P <0.05), but there was no obvious variation of Bax and Fas expression( P >0.05). Inhibition of proliferation and apoptosis were not obvious in PML-RARα negative HL-60 cells induced by low concentration As2O3 ( P >0.05), and there was no obvious variation of Bcl-2, Bax, Fas mRNA expres-sion or Bcl/Bax ratio( P >0.05). Conclusion The ratio of Bcl-2/Bax is contributed to the different sensitiv-ities of PML-RARα negative HL-60 cells and positive NB4 cells induced by low concentration of As2O3.
5.Preliminary curative effect of interspinous blocking injection guiding by X-ray for the diagnosis and treatment of lumbar Baastrup's disease.
Kan LIU ; Wen-Wen WU ; Ji-Dong GUO ; Dong-Feng REN ; Li LI ; Hua-Dong WANG ; Jia TANG
China Journal of Orthopaedics and Traumatology 2014;27(3):220-223
OBJECTIVETo evaluate the preliminary curative effect of interspinous injections for the diagnosis and treatment of back pain caused by lumbar kissing spine (Baastrup's disease) under fluoroscopically guiding.
METHODSFrom November 2011 to March 2013,17 patients with back pain caused by Baastrup's disease were treated with fluoroscopically-guided interspinous injections, including 7 males and 10 females with an average age of 49.6 years old ranging from 40 to 71 years old; the duration of the disease ranged from 2 to 5 years with a mean of 3.7 years. The visual analogue scale (VAS) and the lumbar segments range of motion (ROM) was analyzed at pre-operation, 2 days, 3 months and final followed-up after operation, the effects were evaluated with modified Macnab standard.
RESULTSAll patients were follow-up from 6 to 10 months with an average of 7.6 months. The pre-operative VAS was 6.41 +/- 0.94, the postoperative VAS at different time points improved significantly comparing with pre-operation,and the differences were statistically significant (P < 0.01). There was no significant difference in VAS at different time points after operation (P > 0.05). The ROM of operated segment and adjacent segment was (4.88 +/- 0.86) degrees and (6.82 +/- 0.73) degrees respectively at pre-operation. The postoperative operated segment ROM at different time points improved significantly comparing with pre-operation, and the differences were statistically significant (P < 0.05). Compared with pre-operation, there was no significant difference in adjacent segment ROM at different time points after operation (P > 0.05). According to modified Macnab, the result was excellent in 6 cases, good in 7 cases, fair in 3 cases and poor in 1 case.
CONCLUSIONFluoroscopically-guided interspinous injections is an effective method for the diagnosis and treatment of Baastrup's disease. The method has advantages of simple operation, minimal invasion and safety, satisfactory short-term and medium-term therapeutic effect; it can also effectively lessen the pain of lumbar and back.
Adult ; Aged ; Anesthetics, Local ; administration & dosage ; Female ; Humans ; Low Back Pain ; diagnosis ; diagnostic imaging ; drug therapy ; Lumbar Vertebrae ; diagnostic imaging ; drug effects ; Male ; Middle Aged ; Nerve Block ; Spinal Diseases ; diagnosis ; diagnostic imaging ; drug therapy ; Tomography, X-Ray Computed
6.Potential factors associated with clinical stage of nasopharyngeal carcinoma at diagnosis:a case–control study
Ren JUN-TING ; Li MENG-YU ; Wang XIAO-WEN ; Xue WEN-QIONG ; Ren ZE-FANG ; Jia WEI-HUA
Chinese Journal of Cancer 2017;36(10):478-487
Background: In China, most patients with nasopharyngeal carcinoma (NPC) are diagnosed at a late stage and con-sequently have a poor prognosis. This study aimed to investigate potential factors associated with the clinical stage of NPC at diagnosis. Methods: Data were obtained from 118 patients with early-stage NPC and 274 with late-stage NPC who were treated at Sun Yat-sen University Cancer Center between August 2014 and July 2015. Patients were individually matched by age, sex, and residence, and a conditional logistic regression model was applied to assess the associa-tions of clinical stage at diagnosis with socioeconomic status indicators, knowledge of NPC, physical examinations, patient interval, and risk factors for NPC. Results: Although knowledge of early NPC symptoms, smoking cessation, and patient interval were important fac-tors, the number of cigarettes smoked per day, motorbike ownership, and physical examination exhibited the strong-est associations with the clinical stage of NPC at diagnosis. Compared with smoking fewer than ten cigarettes a day, smoking 10–30 cigarettes [odds ratio (OR) 4.03; 95% confidence interval (CI) 1.11–14.68] or more than 30 cigarettes (OR 11.46; 95% CI 1.26–103.91) was associated with an increased risk of late diagnosis. Compared with not owning a motorbike, owning a motorbike (OR 0.38; 95% CI 0.23–0.64) was associated with early diagnosis. Subjects who under-went physical examinations were less likely to receive a late diagnosis than those who did not undergo examinations (OR 0.50; 95% CI 0.28–0.89). However, indicators of wealth were not significant factors. Conclusions: Initiatives to improve NPC patient prognosis should aim to promote knowledge about early symptoms and detection, health awareness, and accessibility to health facilities among all patients, regardless of socioeconomic status.
7.Determination of phillyrin and forsythoside A in Lianqiao Bingdu Qing capsule by RP-HPLC.
Wen-Hua ZHAO ; Ren-Bing SHI ; Bin LIU ; Lan-Zhen ZHANG
China Journal of Chinese Materia Medica 2005;30(1):36-39
OBJECTIVETo establish methods for the determination of phillyrin and forsythoside A in Lianqiao Bingdu Qing capsule by RP-HPLC.
METHODThe determination of phillyrin was carried out on YWG-C18 column (4.6 mm x 250 mm, 10 microm), using acetonitrile-water (25:75) as mobile phase at a flow rate of 0.8 mL x min(-1) and detected at the wavelength 277 nm. The determination of forsythoside A was carried out with YWG-C18 column(4.6 mm x 250 mm,10 microm), using acetonitrile-water-Acetic acid (17:83:0.4) as mobile phase at a flow rate of 1.0 mL x min(-1) and detected at the wavelength 280 nm.
RESULTThe average recovery of phillyrin was 99.6%, RSD = 1.9% (n = 5). The average recovery of forsythoside A was 101.3%, RSD = 2.5% ( n = 5).
CONCLUSIONThe methods were simple and accurate and could be used to control the quality of the Lianqiao Bingdu Qing capsule.
Capsules ; Chromatography, High Pressure Liquid ; methods ; Drugs, Chinese Herbal ; administration & dosage ; chemistry ; isolation & purification ; Forsythia ; chemistry ; Fruit ; chemistry ; Glucosides ; analysis ; Glycosides ; analysis ; Plants, Medicinal ; chemistry
8.Growth of breast cancer cells is inhibited in vitro by cyclin E-siRNA.
Wen-shan HE ; Tao HUANG ; Jing-hua REN
Chinese Journal of Oncology 2009;31(3):170-173
OBJECTIVETo investigate the changes of biological characteristics of breast cancer cell line by cyclin E expression.
METHODSHuman breast cancer cell line MCF-7 was transfected with cyclin E siRNA vector pEGFP/CCNE2. siRNA-induced silencing of cyclin E was determined by RT-PCR at RNA level and Western blot at protein level. The proliferation of MCF-7 cells and their sensitivity to chemotherapy was measured by CCK-8 assay. The cells were examined by FCM. The cell line was injected into nude mice and the tumor size was measured.
RESULTSThe expression of cyclin E was inhibited in the MCF-7 cells. The relative expression level of cyclin E mRNA was 0.23 +/- 0.05, and that of cyclin E protein was 0.24 +/- 0.05. The cell growth was inhibited by 68.56% +/- 0.08%, and their sensitivity to chemotherapy was increased. Most cells were blocked at G(1) (77.38%), their tumorigenic ability in nude mice was reduced, and the size of tumor formed in mice of the experimental group was decreased than that of controls.
CONCLUSIONInhibition of cyclin E expression in breast cancer cells can block their cell cycle at G(1) phase, reduce their cell growth, differentiation and proliferation, and increase their sensitivity to chemotherapy.
Animals ; Antibiotics, Antineoplastic ; pharmacology ; Antimetabolites, Antineoplastic ; pharmacology ; Antineoplastic Agents, Phytogenic ; pharmacology ; Breast Neoplasms ; metabolism ; pathology ; Cell Cycle ; Cell Line, Tumor ; Cell Proliferation ; drug effects ; Cyclin E ; genetics ; metabolism ; Doxorubicin ; pharmacology ; Female ; Fluorouracil ; pharmacology ; Genetic Vectors ; Humans ; Male ; Mice ; Mice, Inbred ICR ; Mice, Nude ; Neoplasm Transplantation ; Paclitaxel ; pharmacology ; Plasmids ; RNA Interference ; RNA, Messenger ; metabolism ; RNA, Small Interfering ; genetics ; Transfection ; Tumor Burden
9.Down-regulation of Nrf2 enhances the cytotoxicity of Oxaliplatin to H460 cells.
Ai XIN ; Hua-yu REN ; Xiu-wen TANG
Journal of Zhejiang University. Medical sciences 2010;39(1):11-16
OBJECTIVETo establish a stable H460 cell line with Nrf2 down-regulation to study the role of Nrf2 in Oxaliplatin resistance.
METHODSNrf2 down-regulated H460 cell line was obtained by transfecting cells with pRS-hNrf2 and followed by screening for Nrf2 expression by Western blotting. The cell lines were further characterized by analysing cellular GSH levels and cytotoxicity to anti-cancer drugs with MTS.
RESULTNrf2 down-regulated H460 cell lines were established successfully. Cellular GSH level reduced significantly in the H460-C9 and H460-C13 compared to control cells (P(C9)= 0.00249, P(C13)= 0.03944). Down-regulation of Nrf2 in H460 sensitized the anti-cancer effect of Oxaliplatin and Doxorubicin.
CONCLUSIONNrf2 plays an important role in drug resistance. Down regulation of Nrf2 in H460 cell enhances cytotoxicity of Oxaliplatin.
Antineoplastic Agents ; pharmacology ; Carcinoma, Non-Small-Cell Lung ; genetics ; metabolism ; pathology ; Cell Line, Tumor ; Cell Proliferation ; drug effects ; Down-Regulation ; Drug Resistance, Neoplasm ; genetics ; Humans ; Lung Neoplasms ; genetics ; metabolism ; pathology ; NF-E2-Related Factor 2 ; genetics ; metabolism ; Organoplatinum Compounds ; pharmacology ; Transfection
10.The association of insulin receptor substrate 2 gene polymorphism with type 2 diabetes and its related metabolism
Li-Lin GONG ; Su-Hua ZHANG ; Rong LI ; Wei REN ; Zeng-Chan WANG ; Xiao-Su BAI ; Wen-Yu ZHANG ;
Chinese Journal of Endocrinology and Metabolism 2000;0(06):-
Objective To study the genotype distribution of insulin receptor substrate-2(IRS-2)gene 1057G/A polymorphism in Han population from Southwest China,and to explore its association with the metabolism of glucose and lipids,insulin resistance and islet?-cell function in type 2 diabetic patients and subjects with impaired glucose tolerance(IGT).Methods A total of 929 Hans[462 patients with type 2 diabetes(DM group) and 164 subjects with IGT(IGT group)and 303 normal controls(NC group)]from Chongqing and nearby regions were screened for 1057G/A polymorphism of IRS-2 gene by PCR-RFLP assay.Body mass index(BMI),plasma glucose,serum insulin and lipid profile,high-sensitive C-reactive protein(hsCRP)and non-esterified fatty acid were measured.Homeostasis model assessment of insulin resistance(HOMA-IR)and disposition index(DI)were used to estimate insulin resistance and?-cell function respectively.Results In DM group,A allele frequency was significantly lower than that in NC group(0.326 vs 0.388,X~2=6.19,P=0.01).Compared with NC group,AA genotype frequeney was lower and GG genotype frequeney was higher in DM group(0.104 vs 0.135 and 0.452 vs 0.360 respectively,X~2=6.80,P