The characteristics of EPPB41L3 methylation in esophageal squamous cell carcinoma tissues and in plasma and the clinical significance of this methylation / 实用医学杂志

Objective To detect EPPB41L3 methylation frequency difference between esophageal squamous cell carcinoma (ESCC) tissues and the normal tissues and between ESCC patients′plasma and healthy volenteers′plasma, and to analyze the correlation with clinicopathological parameters. Methods We collected esophageal squamous cell carcinoma tissues (n = 42 patients) and adjacent surrounding normal tissues (n = 42 patients), and plasma from 42 patients with ESCC and from 50 healthy individuals. We used methylation specific PCR (MSP) combined with agarose gel electrophoresis to detect the methylation status of the EPB41L3. We used the SPSS 13.0 software for statistical analysis by χ2 test and Fisher′s exact test. Results EPB41L3 frequency of methylation was significantly higher in tumor tissues than in the adjacent tissues (59.5% vs. 4.8%), the difference was statistically significant (χ2 = 28.873, P < 0.001). For plasma, EPB41L3 methylation frequency was 31.0%in cancer patients, while was not detectable in the healthy volunteers. Methylation of EPB41L3 in tissues was more frequently found in patients with tumor size of ≥ 5 cm or T3 than in patients with tumor size of < 5 cm or T1-2. Conclusions The methylation frequency of EPB41L3 is higher in ESCC tissues than in control normal tissues, and higer in plasma from ESCC patients than that from the healthy volunteers. EPB41L3 methylation is more frequently found in patients with more advanced disease.

Intratumor injection of recombinant attenuated salmonella carrying Mycobacterium tuberculosis heat shock protein 70 and herpes simplex virus thymidine kinase genes to suppress murine melanoma growth / 南方医科大学学报

<p><b>OBJECTIVE</b>To study the effection of suppression murine melanoma growth by Intratumor injection of recombinant attenuated salmonella carrying heat shock protein 70 and herpes simplex virus thymidine kinase genes.</p><p><b>METHODS</b>Plasmids PCMV-mtHSP70-IRES-TK were electro-transferred into salmonella typhimurium SL7207 to construct recombinant salmonella typhimurium. In vivo, Recombinant bacteria were injected into the mouse melanoma and the antitumor effection was observed. The survival period was recorded and safety analysis for this vaccine in each group.</p><p><b>RESULTS</b>In vivo, the mtHSP70/HSV-tk recombinant bacteria can suppress tumor growth significantly and extend survival. After recombinant Salmonella, 10(9) CFU/mL, was administered as an intratumoral injection, No diarrhea were observed. During therapy, body weight did not change markedly.</p><p><b>CONCLUSION</b>Results of the animal experiment suggests intratumor injection of recombinant attenuated salmonella typhimurium containing mtHSP70 and HSV-tk genes, has targeting ability against B16 tumor cell and could significantly inhibit tumor growth .</p>

Expression and cytotoxic effect of transmembrane form of human blood group A antigen mimotope vaccine in a malignant melanoma cell line B16 / 中华皮肤科杂志

Objective To establish a stable cell line expressing transmembrane form of human blood group A antigen mimotope vaccine by transfecting malignant melanoma cell line B16, and to detect the cytotoxicity of the vaccine against melanoma cells. Methods Cultured B16 cells were classified into 4 groups, i.e.,P/F-M-pIRES group [transfected with the recombinant plasmid mimotope peptide/Fas-macrophage inflammatory protein (Mip)-pIRES], P/F-pIRES group (transfected with the recombinant plasmid mimotope peptide/FaspIRES), M-pIRES group (transfected with the recombinant plasmid Mip-pIRES), and pIRES group (transfected with the empty plasmid pIRES). B 16 cells were transfected through Lipofectamine 2000. Subsequently, RT-PCR and Western blotting were performed to detect the mRNA and protein expressions of the mimotope peptide/Fas fusion gene and Mip3β in transfected B16 cells. Cell counting kit-8 (CCK-8) was used to evaluate the vaccinemediated complement dependent cytotoxicity (CDC) and antibody-dependent cell-mediated cytotoxicity (ADCC)against B16 cells. Results RT-PCR yielded specific DNA fragments with expected size. Western blotting revealed the anti-A antibody-binding activity of the recombinant mimotope peptide/Fas fusion protein. Factor analysis indicated significant differences in CDC (F = 244.522, P ＜ 0.01 ) and ADCC (F = 71.593, P ＜ 0.01 )against B16 cells between the 4 groups. Group comparisons demonstrated more intense CDC and ADCC in P/FM-pIRES and P/F-pIRES groups compared with M-plRES and pIRES groups, stronger ADCC in P/F-M-pIRES group in comparison with P/F-pIRES group (F = 15.42, P ＜ 0.05), but no significant difference in CDC was observed between M-pIRES and pIRES group. Conclusions The transmembrane form of human blood group A antigen mimotope vaccine could be stably expressed in B16 cells, and mediate ADCC and CDC against B16 cells in vitro.

Research on the apoptosis of malignant melanoma cell induced by transmembrane form of human blood group A mimotope vaccine / 中国医师杂志

Objective To investigate the apoptotic effect of the transmembrane form vaccine of human blood group A mimotope on malignant melanoma cell line B16. Methods B16 cells were transfected with different recombinant plasmid through Lipofectamine 2000 and incubated with different concentration of monoclonal anti-A antibody at 2.5 μg/ml, 5 μg/ml,10 μg/ml and 20 μg/ml. Apoptosis rate of cells was determined with Annexin Ⅴ/PI double staining by flow cytometry. Results Apoptosis rate to P/F-M-pIRES group B16 cells was 74.74% when anti-A monoclonal antibody concentration was 10 μg/ml; apoptosis rate of plasmids carrying peptide/Fas fusion gene such as P/F-M-pIRES group and P/F-pIRES group were significantly higher than M-pIRES group and pIRES group. The apoptosis rate was statistically significantly different between different recombinated plasmid groups (F＝669.707,P＜0.01). The apoptosis rate was statistically significantly different between different antibody groups (F＝106.596,P＜0.01). The interaction between recombinated plasmid groups and antibody groups was statistically significant (F＝34.806,P＜0.01). Conclusions The transmembrane form vaccine of human blood group A mimotope could induce B16 cell apoptosis in vitro. This vaccine may be a promising candidate for potential malignant melanoma therapy.

Effect of L-gossypol on apoptosis in human nasopharyngeal carcinoma cell / 中国基层医药

Objective To investigate the apoptosis-inducing effect of L-gossypol on human nasopharyngesl carcinoma cell line CNE2 and its possible mechanism.Methods The effect of L-gossypol on proliferation of CNE2 cells was estimated by MTT assay.Flow cytometry was used to analyz cell apoptosis induced by(-)-gossypol and the expression of Bcl-2 、Bax proteins.Caspase-3 activity was determined by caspase-3 colorimetric assay kit.Results Lgossypol was able to inhibit the proliferation of CNE2 cells in a dose-dependent and time-dependent fashion at concentrations more than 10 μmoL/L.L-gossypol regulated cell cycle and GO/G1 arrest could be observed in CNE2 cells.In the process of apoptosis,the expression of Bcl-2 was down-regulated and Bax was up-regulated,caspase-3 activity was in peak at 24h.Conclusion L-gossypol could induce apoptosis in nasopharyngeal carcinoma cell line CNE2 in vitro,which may be associated with down-regulation of Bcl-2,up-regulation of Bax genes expression and caspase-3 activation.

Construction of tissue-engineered artificial nerve with the compound of acellular nerve graft and bone marrow mesenchymal stem cells to treat sciatic nerve defect / 中国组织工程研究

BACKGROUND: Tissue-engineered artificial nerve was successfully constructed with the compound of acellular nerve graft and bone marrow mesenchymal stem cells, suggesting that it could promote peripheral neural regeneration.OBJECTIVE: To construct tissue-engineered artificial nerve, and to verify neural functional recovery of bridging rats following sciatic nerve defect.METHODS: A total of 60 adult male SD rats were used to induce sciatic nerve defect models (15 mm in length), and they were then randomly divided into three groups, with 20 rats in each group. Sciatic nerve defect group was treated with tissue-engineered artificial nerve; blank control group was treated with tissue-engineered nerve stent; autoallergic neural control group was treated with autoallergic neural transplantation. Twelve weeks after bridging, histology of sciatic nerve and neuralfunctional recovery were detected via gross observation, wet mass of tibialis anterior muscle, and histological analysis.RESULTS AND CONCLUSION: At 12 weeks after bridging surgery, rats in experimental group were able to stand on the floor,and withdrawal reflex was detected at plantar skin on the surgical side. S-100 protein of plantar skin was positive. There was no significant difference in wet mass of tibialis anterior muscle between experimental and autoallergic neural transplantation group (P ＞ 0.05). HRP retrograde tracing in the experimental group demonstrated that HRP-positive cells were observed in both spinalcord and posterior root ganglion. There was no significant difference in number of myetinated nerve fiber, thickness of myelin sheath, and area of nerve tissue between experimental and autoallergic neural transplantation group. The results demonstrated that the compound of acellular nerve graft and bone marrow mesenchymal stem cells could successfully construct tissue-engineered artificial nerve to repair sciatic nerve defect and promote neurohistological reconstruction and functional recovery.

Antitumor efficacy of intratumoral injection of xenoantigen in mice / 中国组织工程研究

BACKGROUND: Herterologous antigen has strong immunogenicity and easily induces immunological response. Introduction of herterologous antigen into tumor may induce a serial of immunological reactions in the tumor and may reverse the immunosuppression of tumor microenvironment to treat tumor.OBJECTIVE: To evaluate the antitumor efficacy of intratumoral injection of human erythrocyte membrane antigens in micebearing S180 sarcoma.METHODS: Kunming mice bearing S180 sarcoma model were established and treated with 5 g/L human erythrocyte membrane antigens suspension or normal saline for five days. Tumor volume was calculated before the first injection and 3, 7, and 14 days after the first injection. In addition, the tumor cells in combination with human erythrocyte membrane antigens group, the njectionof saline group (the control group), and the injection of human erythrocyte membrane antigens or saline group (pre-immunized by suspension of human erythrocyte of blood group type A). Another 60 mice bearing S180 sarcoma were established and subjected to the above pre-immunization and injection of saline or human erythrocyte membrane antigens. Six mice selected from each group were sacrificed 14 days after the first injection, and tumors were weighed, followed by histological examination. Survival of remainders in each group was observed.RESULTS AND CONCLUSION: Tumor volumes in each group increased gradually. Tumor volumes in the human erythrocyte membrane antigens injection group, the tumor cells in combination with human erythrocyte membrane antigens group, and the human erythrocyte membrane antigens injection group (immunized) were smaller than the control group, Intratumoral injection of human erythrocyte membrane antigens significantly reduced tumor weights. Tumor necrosis, infiltration of inflammatory cells such as lymphocytes were observed in tumor tissues section examination following the intratumoral injection of human erythrocyte membrane antigens. The mouse survival time showed no statistical difference among different groups. Intratumoral injection of heteroloaous ervthrocvte membrane antiaens can inhibit tumor arowth of S180 sarcoma bearina mice.

Repairing rat sciatic nerve defects with the compound of acellular nerve graft and bone marrow mesenchymal stem cells / 中国组织工程研究

BACKGROUND: Previous studies have successfully prepared the natural and biologically degraded acellular nerve graft and have proved the effect of promoting neural regeneration.OBJECTIVE: To construct tissue engineered artificial nerve with acellular nerve graft and bone marrow mesenchymal stem cells, and to observe the effect of promoting motor functional recovery and repairing rat sciatic nerve defects. DESIGN, TIME AND SETTING: Randomized control animal experiment was performed in the Medical TIssue Engineering Laboratory of the First Affiliated Hospital of Liaoning Medical University between June 2008 and February 2009. MATERIALS: Wistar adult healthy male rats weighing 180-200 g were used to prepare acellular nerve graft, while Wistar adult healthy male rats weighing 100-120 g were used to prepare bone marrow mesenchymal stem cells. Tissue engineered artificial nerve was produced with acellular nerve graft co-cultured with bone marrow mesenchymal stem cells. METHODS: Sixty Wistar adult healthy male rats weighing 180-200 g were induced sciatic nerve defect models, 15 mm long. SD rats were divided into three groups at random with 20 animals in each group. ①Experiment group: Rat sciatic nerve defects were bridged with tissue engineered artificial nerve. ②Blank control group: Rat sciatic nerve defects were bridged with tissue engineered nerve scaffold. ③Autologous nerve control group: Rat sciatic nerve defects were bridged with autologous nerve graft. MAIN OUTCOME MEASURES: At 12 weeks postoperation, the recovery of motor function was evaluated with gross observation, electrophysiology, histological observation and triceps surae wet weight.RESULTS: ①At 12 weeks postoperation, the toes at the operation side could separate and supported to the ground in the experiment group; there was no significant difference in the regenerated nerve conduction velocity between experimental group and autologous nerve graft group. ②At 12 weeks postoperation, histochemical stain results showed AchE-positive motor end-plate arranged regulady in the middle and superior part of gestrocnemius muscle to form end-plate zone in the experiment group. By use of silver staining, the regenerated nerve tract and the emergent branch were shown to be connected with motor end-plate.③There was no significant difference in the tibialis anterior muscle wet weight between experimental group and autologous nerve graft group. CONCLUSION: Bridging acellular nerve graft and bone marrow mesenchymal stem cells into rat sciatic nerve defects can promote motor functional recovery.

Influence of infrasound therapy on Raji cells / 中华物理医学与康复杂志

Objective To investigate the influence of infasound therapy on Raji cells. Methods The Raji cell line was cultivated routinely and grouped as an infrasound and a control groups.Infrasound 8TM was used as a therapeutic infrasound generator which worked in 3 modes(frequency range 4～20 Hz,infrasound energy less than 90 dB).The applicator of the infasound generator was put on 1.5～2.0 cm fom the surface of liquid in the dish containing Raji ceil.Raji cells would be treated for 15,30,60,and 90 minutes,then tested with trypan blue assay,MTT assay,flow cytometry anatysis,and scanning electron microscope(SEM)after 0,24,and 48 h cultivations,respectively. Results Trypan blue assay showed that there were no significant differences between the 2 groups(P>0.05).MTT assay showed that although optical density value of the infrasound groups seemed to be lower than that of the control group.the differences were not significant(P>0.05).Flow cytometry analysis showed that the rate of necrotic cells and apoptosis cells in all groups was less than 10%;and that the differences between all groups were not significant(P>0.05).The scanning electron microscopy showed that the cells treated by infasound exposure for 120 minutes and then cuhivated for 24 h showed that the prominent or micro-floss of the membrane become shorten and decreased.The surface of the membrane became smooth. Conclusion Infrasound(less than 90 dB)treatment in the experiment had no obvious influence on multiplication and apoptosis of Raji cells.But the membrane of Raji cell Would be affected directly by the infrasound,and the penetration of the membrane could be changed.

Experimental study of rat sciatic coagulation with a novel microwave antenna / 中华物理医学与康复杂志

Objective To observe the sequential ultrastroctural and electrophysiological changes in the sciatic nerve coagulated by a newly-designed microwave antenna. Methods A total of 75 Sprague-Dawley rats were randomly divided into groups A,B and C and irradiated with microwaves at 10,20 or 30 Watts,for 6 seconds to coagulate the left sciatic nerve.Electrophysiological effects and sequential uhrastructural changes were observed on the 0th,2nd,7th,30th and 60th days after coagulation.A static sciatic index was calculated based on measurements of the footprint on the 7th,30thand 60th days after coagulation.Results On the Oth,2nd,7th and 30th days after cpagulation,the static sciatic index,the nerve conduction velocity and the amplitude of the action potentials in groups B and C had decreased significantly compared with those before coagulation.On the 60th day after coagulation.significant recovery was observed in groups A and B,but not in group C.Only mild alteration in uhrastructure was found,and only in group A.The prominent changes in uhrastructure in group B included broken Schwann cell membranes and myelin disintegration.There were severe injuries in group C,including myelin disintegration,cell deformity,coagulative necrosis,axon necrosis,basement membrane necrosis and demyelination.The structure of the sciatic nerve in group B had partially recovered after 60 days,but group C showed no recovery at all. Conclusion Microwave coagulation of a nerve can block its conduction.and even destroy the nerve.Percutaneous microwave coagulation is clinically feasible and call be an alternative treatment for pain.

Rat skeletal muscle alterations after coagulation with novel microwave antenna / 中国组织工程研究

AIM: To evaluate the feasibility and safety of microwave coagulation with novel microwave antenna through observing physiological and pathological changes after the coagulation on rat skeletal muscle. METHODS: The experiment was performed at the Central Laboratory of Zhujiang Hospital of Southern Medical University from December 2006 and September 2007. ①Using MTC-3C microwave tissue coagulation device, monopole microwave antenna after anti-adhesion was vertically inserted into SD rat skeletal muscle for heat coagulation at 20 W?8 s. ②The instant enzyme activity changes after microwave coagulation therapy (MCT) was detected by 2, 3, 5-Triphenyl Tetrazolium Chloride (TTC) staining. The cell apoptosis rate in outer fields of coagulation was evaluated by flow cytometry 0, 12 and 24 hours after coagulation, and the treated muscle was examined pathologically on days 0, 2, 7, 30, and 60. RESULTS: ①The coagulation areas were stained in white immediately after MCT by TTC staining, while the outer normal areas were in red. There were apoptotic cells around coagulation areas, and the highest percentage appeared 12 hours later, reduced thereafter. ②In 24 hours after coagulation, only mild alteration of cellular morphology was found. Two days after MCT, necrosis cells were prominent in transition zone, and the inflammation cells were predominantly composed of macrophage and lymphocytes in the zone, while the cells in the coagulation zone remained structure; 7 days after MCT, the fibroblast and granulation tissue occurred in the transition zone, and the myocyte in the coagulation zone were of necrosis; 30 days after MCT, there were still many inflammation cells in the coagulation zone with some collagenous fibrils in the transition zone; 60 days after MCT, the necrosis cells reduced and the outer margin of the transition zone was encapsulated by marked collagenous fibrils. Tissue repair started from the outer margin of the transition zone to the central coagulation zone. CONCLUSION: Novel microwave tissue coagulation is safe and feasible to be used in the treatment of some minor-carcinoma and some tumors on body face.

Study for preparation and anticancer activity of the stealth epirubicin chitosan nanoparticles / 基础医学与临床

Objective To prepare a new type of chitosans nanoparticles(PEG/CS-EPI NPs),which contains epirubicin and modified by PEG.Furthermore,to investigate the anticancer activity of the NPs in vitro and in vivo.Methods The ionic gelation technique was employed to prepare the PEG/CS-EPI NPs and CS-EPI NPs.The particle size and shape were illustrated respectively by laser scattering and transmission electron microscopy.The proliferation of nasopharyngeal carcinoma cells was detected by MTT assay;The mouse model of implantation murine sarcoma cells(S-180) was applied to evaluate the anticancer effectiveness of PEG/CS-EPI NPs and CS-EPI NPs in vivo.Results The PEG modified CS NPs were discrete and uniform spheres with average diameter of 322.1 nm.The rate of drug loading and encapsulation is 13% and 74% respectively.The results of the anticancer tests showed a sustained cytotoxicity of loading drug NPs on nasopharyngeal carcinoma cells in vitro and the stealth nanoparticles was more powerful than ordinary nanoparticles on the inhibitory potency in vivo.Conclusion Stealth chitosan nanoparticles as compared with ordinary chitosan nanoparticles seems to be a potential candidate of chemotherapy drug carriers.

Effects of anti-HPV16E6-ribozyme on phenotype and gene expression of a cervical cancer cell line / 中华医学杂志(英文版)

<p><b>OBJECTIVE</b>To investigate the effects of anti-HPV16E6-ribozyme (HRz) on phenotype and gene expression of a cervical cancer cell line.</p><p><b>METHODS</b>HRz was designed by computer programs. HRz's activity was identified by cleavage experiments in vitro. HRz and empty eukaryotic plasmids were transfected into CaSKi cells with lipofectin, then renamed CaSKi-R and CaSKi-P, respectively. The expression of ribozyme in transfected cells was observed by RNA dot blot. The amounts of E6 mRNA in three kinds of cells lines were detected by Northern blot. Cell growth curves and soft agar forming ability were studied. The ability of each cell line to form tumors was assessed in nude mice. Apoptosis rates and expression of c-myc, bcl-2, p53 and Fas were detected by flow cytometry (FCM). Antigens of tumor cells, HLA-1, HLA-2, B7-1 and B7-2 were also detected. NK, LAK, and CD(3)AK cells were induced. Their cytotoxicities were detected in CaSKi-R, CaSKi-P, and CaSKi cells.</p><p><b>RESULTS</b>In vitro cleavage reaction demonstrated that HRz could cleave HPV16E6 mRNA in a site-specific manner. HRz could be expressed stably in transfected CaSKi cells. Northern blot analysis showed that E6 mRNA levels were lower in CaSKi-R than in CaSKi. The growth rate of CaSKi-R was slower than those of CaSKi and CaSKi-P. The soft agar-forming rate of CaSKi-R was lower compared with those of CaSKi and CaSKi-P cells. The ability of CaSKi-R to form tumors in nude mice was also poor. The apoptosis rate of CaSKi-R cells was much higher than those of CaSKi and CaSKi-P. HRz could reduce the expression of E6, c-myc and bcl-2 proteins, and increase the expression of p53 as well. HRz could increase the expression of HLA-2, B7-1 and B7-2 antigens. The cytotoxicity of NK, LAK and CD(3)AK cells was much higher in CaSKi-R than in CaSKi-P and CaSKi cells.</p><p><b>CONCLUSION</b>HRz not only reverses the malignant phenotype of CaSKi cells partially, but also induces apoptosis in the cells, and increases sensitivity of CaSKi cells to immune cells.</p>

INVESTIGATION ON TUMOR ASSOCIATED NEUTRAL GLYCOSPHINGOLIPIDS / 解放军医学杂志

To investigate the roles of neutral glycosphingolipids (N-GSLs) in the development of tumor, tumor antigen expression and immunological evasion, the regularities of the expression of N-GSLs in human embryonic and neoplastic tissue were studied, the influence of N-GSLs in the growth regulation, immunological evasion and multidrug resistance(MDR) of tumor cells were analyzed. The tumor embryonic associated antigen CDH, tumor multidrug resistance associated N-GSLs CMH and neoplasm inhi-bitor globoside were found. The significance of N-GSLs synthesis inhibition and de-N-glycosylation in the reversion of MDR and tumor therapy was also partly disclosed.

PRELIMINARY STUDY ON ANTITUMOR ACTIVITY OF A NEW ANTHRACYCLINE: R_5 / 解放军医学杂志

R 5 was administered in vitro to observe its antitumor activity in human breast carcinoma cell line MCF 7. As the most useful and efficient anthracycline, epirubicin(EPI) was served as the positive chemical treatment control. MTT colorimetric assay was applied to detect cytotoxicity of R 5 to MCF 7 cells. Apoptotic rate of cells double marked with Annexin V FITC and PI was examined by flow cytometry. And, the alteration of wild type (WT) p53, bax and bcl 2 proteins expression was also observed. Ultrastructural change of MCF 7 cells was observed under a transmission electron microscope. The results showed that growth restrain was observed in MCF 7 cells when administered with different doses of R 5 or EPI. The effect was increased concomitantly with the increasing of R 5 or EPI concentration and culture time. Apoptosis was observed since 6 hours after the MCF 7 cells cultured with R 5 or EPI, and the effect was increased with the culture time extending and reached the highest peak at about 48 hours. However, the apoptotic rate decreased when cultured for 72 hours. Different doses of R 5 and EPI can all induce apoptosis in MCF 7 cell, and the apoptotic rate increased with their concentration, but decreased when the concentration was higher than 5?10 -6 mol/L. Ultrastructure of apoptotic MCF 7 cells, observed by transmission electron microscope, showed typical morphologic changes of apoptosis.

Modulation of multidrug resistance by 1-phenyl-2-palmitoylamino-3-morpholino-1-propanol in SKOV3-adriamycin-resistant cell line / 中华妇产科杂志

Objective To study the modulation of mdr1 and P glycoprotein (P gp) by 1 phenyl 2 palmitoylamino 3 morpholino 1 propanol (PPMP) in SKOV3 adriamycin resistant (SKOV3/AdrR) cell line Methods SKOV3/AdrR cells were treated with PPMP, mRNA expression of multidrug resistant (mdr1) gene was analyzed by reverse transcriptase polymerase chain reaction Intracellular rhodamine (Rh123) concentration was measured by flow cytometry Results PPMP was found to inhibit mdr1 expression of SKOV3/AdrR at the mRNA level This modulation of gene expression was content dependent and complete inhibition appeared at 25 ?mol/L PPMP treatment PPMP could increase intracellular Rh123 accumulation in SKOV3/AdrR cells After 15, 25 ?mol/L PPMP treatment, Rh123 accumulation in SKOV3/AdrR was markedly enhanced Rh123 fluorescence intensity were 389 98,426 08 respectively ( P

MODULATION OF MDR1 mRNA EXPRESSION AND REVERSION OF MULTI-DRUG RESISTANCE BY PPMP IN KBv200 CELL LINE / 解放军医学杂志

The current study was designed to investigate the effects of PPMP (DL-threo-1-phenyl-2- palmitoylamino-3-morpholino-1-propanol), a kind of glycolipids synthase inhibitor, on the modulation of mdr1 mRNA expression and the reversing effect of multi-drug resistance by PPMP in human malignancy KBv200cell line. In vitro KBv200 cells were treated with PPMP in different concentration, the alterations of mRNA expression of drug-resistant gene mdr1 in KB (sensitive cell line) and KBv200 (before and after the treatment of PPMP) cells were analyzed by RT-PCR. Intracellular rhodamine(Rh123) concentration was measured by flow cytometry. PPMP was found to inhibit mdr1 gene expression of KBv200 at the mRNA level, and complete inhibition appeared at 25μmol/L PPMP treatment for 48h. PPMP could increase intracellular Rh123 accumulation in resistant cell lines. This modulation of gene expression and Rh123 accumulation was directly correlated with the concentration of PPMP. It suggested that PPMP, a chemical inhibitor of glycolipids synthase, could modulate mdr1 expression at the mRNA level in a content dependent manner, PPMP possesses MDR-reversing activity.

Effects of repeated treatment with different concentrations of chemotherapy drugs on proliferation and chemo-resistance in human lung cancer cell line / 第二军医大学学报

Objective:To explore the variation of tumor cell proteins correlated with cell proliferation and chemo-resis-tance after repeated treatment with chemotherapy drugs.Methods:The human lung cancer cell line(LPET-a-2)was repeat-edly treated with chemotherapy drugs:doxorubicin,etoposide,cisplatin and the combination of the3drugs,and each drug was given at2concentrations.The treatment intervals were recorded.ErbB-2,c-myc,MDR1,MRP,LRP,ref-1and NF-?B were tested by flow cytometry.The cells expressing each protein,the mean and total quantity of each protein after each treat-ment were calculated by different drugs at different concentrations.Results:The levels of every protein decreased along with the time of culture.In high-dose group,every item decreased along with the time of treatment.In low-dose group,there was no rule for the item variation with some decreased and some increased.Conclusion:Low-dose anticancer drugs are easier to induce cell proliferation and chemo-resistance than high-dose one,which suggests that adequate chemotherapy should be given in clinical practice.

Cryotherapy combining with ethanol injection therapy in the treatment of irresec table hepatic cancer / 中国癌症杂志

Purpose:To observe the effect and safety of c om bination therapy with Percutaneous Local Cryotherapy(PLCT)and Percutaneous Etha nol Injection Therapy( PEIT)in patients with irresectable hepatic cancer Methods:43 patients with irresectable hepatic carcinoma were enrolled 29 tumors of 14 patients were treated by PLCT,23 tumors of 16 patients were treated by PEIT,and 23 tumors of 13 patients were treated by combination therapy Results :The tumor size decreased by more than 50% 1 month later in 51 7%(15/29 )of tumors treated by PLCT alone, in 43 5%(10/23)of tumors treated by PEIT alone, 78 3%(18/23)tumors treated by combination therapy The 1-year surv ival rates were 64 3%(9/14) in PLCT group?43 8%(7/16) in PEIT group?8 4 6%(11/13)in combination therapy group The blood levels of AFP decreased by various extents Conclusions:Combination therapy with PLCT a nd PEIT is more effective than PLCT or PEIT alone

Effects of anti-HPV16E6-ribozyme on telomerase activity in cervical carcinoma cells and the related mechanisms / 中国肿瘤生物治疗杂志

Objective: To investigate the effects of HPV16E6-ribozyme on telomerase activity in cervical carcinoma cell line CaSKi and the related mechanisms. Methods: Anti-HPV16E6-ribozyme and blank eucaryotic plasmids were transfected into CaSKi cells via lipofectin, and the resultant cells were named as CaSKi-R and CaSKi-P, respectively. The expression of ribozyme in transfected cells was observed by RNA dot blotting. The expression of E6 mRNA and protein in the 3 kinds of cells were detected by Northern blotting and Western blotting, respectively. Telomerase activity was determined by TRAP-Elisa method; the expression of P53, c-myc, hTERT and hRT mRNA were examined by RT-PCR.Results: RNA dot blotting showed that anti-HPV16E6-ribozyme was stably expressed in transfected CaSKi-R cells. Western blotting showed that the expression of E6 mRNA and protein in CaSKi-R cells was obviously lower than that in CaSKi and CaSKi-P cells. The telomerase activities in CaSKi,CaSKi-P and CaSKi-R cells were (0.89?0.14), (0.90?0.11) and(0.36?0.06),respectively. The inhibitory rate of telomerase activity in CaSKi-R cells was 59.55%, which was significantly lower than those in CaSKi and CaSKi-P cells (P