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Objective To explore the impact of preoperative neoadjuvant chemotherapy on the expressions of multi-drug resistance genes in patients with cervical cancer and its relationship with the effect of chemotherapy. Methods Ninety-eight cervical cancer patients with TP regimen selected to perform preoperative chemotherapy were enrolled in the Affiliated Hospital of Guiyang Medical College between January 2010 and June 2014. Immunohistochemisty (En vision method) was used to determine the expressions of P-gP, GST-π and TopoII of the same patients before and after neoadjuvant chemotherapy and explore the relationship with the effect of chemotherapy. Results The positive expression rates of P-gp and GST-π were 71.43% and 64.29% before chemotherapy and 80.61%and 74.49%after chemotherapy, respectively. The former two had significant differences (P 0.05). Before neoadjuvant chemotherapy, the positive expression of GST-π in the ineffective group was statistically higher than that in the effective group (P 0.05). There was significant correlation in the expressions of P-gp, GST-π and TopoⅡ(P < 0.05) before and after neoadjuvant chemotherapy. Conclusions The expression of P-gp, GST-πand TopoⅡgene may not be affected by the clinical and pathological features of cervical cancer, but may change expressions of multi-drug resistance genes in cervical cancer by neoadjuvant chemotherapy. Monitoring their expression has a guiding significance for drug selection, prognostic judgment, and the following treatment regimen decision. The GST-π, expression level can be used as a biological parameter to predict the effect of TP regimen neoadjuvant chemotherapy.
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Objective To investigate the role of human membrane associated sialidase (Neu3) in multidrug resistance ( MDR) of K562 cells. Methods K562 cells and K562/ADM cells were treated with DNR alone or with combination of DNR and NeuAC2en. The cell survival rate was measured by MTT assay; the expression of MDR related factors and apoptosis related factors were determined by Western blot and RT-PCR; Neu3 activity was detected by TBA reaction. Results The survival rate of K562/ADM cells was higher than that of K562 cells; NeuAC2en showed synergistic effect with DNR(P <0. 01) ; Neu3 activity of K562 and K562/ADM cells decreased after DNR or NeuAC2en induction and the decrease was most significant in the combination treatment group (P <0. 01). Under the identical condition, the protein expression of P-gp and Neu3 and the mRNA expression of MDR1, Neu3, BCL-xl and BCL-2 increased comparing with K562 cells. The mRNA level of BAX in K562 and K562/ADM groupsdid not changed significantly. MDR1, Neu3,BCL-xl and BCL-2 decreased after DNR induction and down-regulated most obviously in the groups treated by DNR combined with NeuAC2en. Conclusion Neu3 may be related with multidrug resistance of K562/ADM cell through regulating apoptosis and the expression of MDR1.
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Objective To discuss the diagnosis of refractory epilepsy (RE) in children, and to study the association of the single nucleotide polymorphisms (SNPs) of muhidrug-resistance gene (MDR1) C3435T with pharmaco- resistant epilepsy. Methods Four hundred children with epilepsy were retrospectively or prospectively identified from multiple sources in our hospital in Shanghai and were followed-up for the occurrence of refractory epilepsy. The clinical features of RE regarding age at onset, gender, seizure type, electroencephalogram, neuroimaging, development of central nervous system, etiology and prognosis etcetera were investigated. DNA samples were obtained from 132 patients with epilepsy (70 RE and 62 responsive epilepsy) and 62 health children by DNA extraction kit. Genotype of the C3435T polymorphism was determined by DNA sequence analysis after traditional polymerase chain reaction. The frequency of genotypes and alleles among the three groups was compared by Chi-square test. Results Eighty-three (20.8%) out of total 400 patients were RE. Among them 65 (78.3%) patients failed at least 2 drugs in six months. Forty-two (50.6%) were administered at least 3 drugs on the last follow-up. Medical treatment showed remarkable effective in 6 (7.2%) RE patients, effective in 40 RE patients (48.2%). No effectiveness was seen in another 37 (44.6%) RE patients, however 25 out of 37 presented symptomatic alleviation. Significant difference in genotype (CC, CT, Tr) frequency was neither found between RE and responsive epilepsy patients nor between RE patients and healthy controls. No association between the C3435T polymorphism in the human MDR1 gene and refractory epilepsy was found by logistic analysis. Conclusions Refractory epilepsy could be diagnosed in 6 months after being treated with anti-epilepsy drugs (AEDs) in children with average attack once per month at least and failed more than 2 AEDs. Multiple AEDs were necessary for treatment. No association between the C3435T polymorphism in the human MDR1 gene and refractory epilepsy was found by logistic analysis in this study.
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Objective To investigate the expression and sensitivity of 6 muhidmg resistance gene products(MDR) : GST-π、LRP、MRP、Topo Ⅱ、P-gp、TS in gastric cancer during neoadjuvant chemotherapy. Methods Expression of the 6 muhidrug resistance gene was detected by immunohistoehemistry in 35 cases of stomach cancer tissues before and after neoadjuvant chemotherapy. The relationship between muhidrug resistance gene and neoadjuvant chemotherapy was analyzed. All patients were given FOLFOX4, and the effects were evaluated according to World Health Organization criteria. Results The overall response rates to chemotherapy was 46%, expression of the 6 mtdtidrug resistance gene in 35 cases of stomach cancer tissues were as follows:GST-π was 40% ,LRP was 69% ,MRP was 34% ,Topo Ⅱ was 37% ,P-gp was 86%, TS was 40%, expression of the 6 muhidrug resistance gene did not change during neoadjuvant chemotherapy; GST-π、LRP、MRP、Topo Ⅱ、P-gp were not correlated with chemotherapy sensitivity(P > 0.05), while TS was significantly correlated with chemotherapy sensitivity (P = 0.0048). Conclusion FOLFOX4 does not effect a change in the expression of the 6 muhidrug resistance gene: GST-π、LRP、MRP、Topo Ⅱ、P-gp and TS, while TS is significantly correlated with gastric cancer chemotherapy sensitivity.
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Objective To explore the effect of gene mutations of arsenic transport proteins-muhidrug resistance-associated proteins(MRP1 and MRP2)on phenotype of endemic arsenic poisoning.Methods Two hundreds and thirty-nine rural residents in 3 villages of Shuocheng Region,Shanxi Province were interviewed and examined by simple random sampling who had been lived there for 20 yearn at least.All the objects were divided into two groups on the basis of clinical examination with"The Standard Diagnosis of Endemic Arsenic Poisoning" (WT/S 211-2001):subjectives with skin lesion as a arsenic poisoning group and without skin lesion as a control group. One hundred and ninety-three blood samples were collected from each participanL Seventy-five arsenic poisoning cases and 118 controls were detected the gene mutations in the 2,17,23 exons of M RPI and the 10,18,31 exons of MRP2 by PCR-single strand conformation polymorphism (PCR-SSCP) and compared by multivariate Logistic regression model. Results Seventy-five cases and 164 controls underwent questionnaires. Age[ (58.85±11.26) vs (45.73±11.92),OR = 3.378,P ＜ 0.05],gender[male,57.3%(43/75)vs 27.4%(45/164),OR = 3.553,P＜ 0.01 ],smoking[46.7%(35/75) vs 21.3%(35/164),OR = 3.225,P ＜ 0.01 ],drinking[ 17.3%(13/75) vs 8.5% (14/164),OR = 1.836,P ＞ 0.05],vegetable and fruit intake[5.3%(4/75) vs 9.1%(15/164),OR = 0.560,P ＞ 0.05],egg and meat intake[34.7%(26/75) vs 30.5%(50/164),OR = 1.210,P ＞ 0.05],exposure of pesticide [41.3%(31/75) vs 29.3%(48/164),OR = 1.864,P ＜ 0.05] were tested by Logistic regression model. There was no gene mutation detected in the 23 exon of MRP1 and the 18 exon of MRP2. The gene mutations frequencies of the 2 exons of MRP1 in arsenic poisoning and control groups were 8.00% (6/75) and 5.93% (7/118),respectively;they were 13.33%(10/75) and 8.47%(10/118) of the 17 exons of MRP1,respectively;they were 22.67%(17/75) and 18.64%(22/118) of the 10 exons of MRP2,respectively;they were 5.33%(4/75) and 2.54%(3/118) of the 31 exons of MRP2,respectively. There was no significant difference between two groups(x2 = 0.312,1.165,0.460, 2.794,respectively,all P ＞ 0.05). After age,gender,smoking,drinking,nutritional level and exposure of pesticide being adjusted by multivariate Logistic regression model,there was no significant difference between two groups (OR = 0.803,1.892,2.388,1.098,respectively,all P ＞ 0.05). Conclusions The gene mutations of 2,17,23 exons of MRPI and the 10,18,31 exons of MRP2 may have no effect on the phenotype of endemic arsenic poisoning.
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Objective To investigate the expression of P-glycoprotein(P-gp), multidrug resistance-associated protein (MRP), lung resistance-related protein (LRP) and predicts treatment outcome in the elderly acute myeloid leukemia. Methods Multi-parameter flow eytometric assay was used to quantify expression of P-go, muhidrug-associated resistance protein (MRP) and lung resistance-related protein (LRP) in bone marrow viable blasts from 12 eldely acute myeloid leukemia patients at diagnosis (M2a:4, one is come from MDS, M3a:2, M5a:5, M6a:1). Correlation of the MDR protein expression with treatment outcome were analysed. Results The frequency of expression of P-gp, MRP and LRP was 58.33 %, 8.33 %, 50 %, respectively and P-gp(+)/MRP(+) 0, P-gp(+)/LRP(+) 33.33 %, MRP(+)/LRP(+) 0, P-gp(+)/MRP(+)/LRP(+)8.33 %, P-go(-)/MRP (-)/LRP(-) 33.33 %;respectively. The frequency of overexpression of P-gp or LRP alone and both of them were relatively higher. The rate of complete remission (CR) and overall survival (OS) at one year of P-gp(+) group were significantly lower than those of P-go(-) group, and LRP(+) group also were lower than LRP(-) group. Conclusion The Frequency of expression of P-gp or LRP and coexpression of both of them were higher in elderly patients with AML. The overexpression of P-gp and/or LRP was a poor prognotic factor for eldely myeloid leukemia in elderly patients.
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Objective To investigate the expression of muhidrug resistance-related protein 1 (MRP1),lung resistance-related protein(LfuP)and glutathione S-transferase-π(GST-π)in carcinoma of the gallbladder and cholangiocarcinoma. Methoils MRP1,LRP,GST-πwere measured in experimental group(18 cases of carcinoma of the gallbladder,36 CSSeS Of cholangiocarcinoma)and control group(13cases of cholecystitis and cholangeitis)by immunohistochemistry.Statistical analysis used chi-square test and spearman test. Results The positive rate of MRP1,LRP,GST-π in carcinoma of the gallbladder and eholangiocarcinoma were 72%(13/18),78%(14/18),61%(11/18)and 86%(31/36),75%(27/36),69%(25/36),respectively,significantly higher than those of 23%(3/13),23%(3/13),23%(3/13)(X2=4.5,P＜0.05)in control group.The expression of LRP[93%(13/14)]in pafients＞60 years old was significantly higher than 64%(14/22)in patients younger than 60 yrs old(x2=3.9,P ＜0.05).In addition,their expression was not related to gender,age,staging,tumor differentiation and lymph node metastasis(P＞0.05).The expression of MRP1 was related with tllose Of GST-π,Spearman correlation coefficient=0.569(P＜0.05).Conclusions MRP1,LRP,GsT-π were over expressed in various degrees in carcinoma Of the gallbladder and cholangiocarcinoma witllout chemotherapy.and related to the primary muhidrug resistance Of cholangiocarcinoma and carcinoma of the gallbladder.
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Objective To establish a multidrug resistance cell line of human bladder tumor and study its characteristics and mechanism of muhidrug resistance. Methods Human bladder tumor cell line T24 was induced to muhidrug resistance cell line T24/ADM by intermittent administration of high dose ADM. The multidrug resistance to multianticancer agents of the ceils was evaluated by MTT assay; the expression of MDR gene was assessed by RT-PCR and P-gp expression was determined by flow cytometry. Results T24/ ADM resisted to many anti-tumor agents, and its IC50 of ADM was 16.3 times higher than that of T24. Significant overexpression of MDR gene and p-gp of the multidrug resistant cells were detected. Conclusion T24/ADM is a human muhidrug-resistant cell line, and it's drug resistance relates to the overexpression of MDR gene and p-gp.
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Aim: To evaluate the effects of HZ08, a novel P-glycoprotein inhibitor, on reversing tumor resistance of K562/ADM to adriamycin in nude mice and on the activities of cytochromes P-450 (GYP) isoforms. Methods: Nude mice bearing K562/ADM were injected at different doses of HZ08 with adriamycin for 4 weeks. The tumor weights of HZ08 treatment groups were determined and compared to those of the control and positive groups. In addition, the effects of HZ08 were examined on GYP isoforms-mediated metabolism of specific substrates by GYP isoforms in rat liver microsomes in the presence or absence of HZ08. Results: The tumor weights of HZ08 treatment groups were significantly decreased and HZ08 was a relatively potent inhibitor of CYP3A4, with no significant effects on other isoforms tested. Conclusion: HZ08 has potent effects on reversing P-glycoprotein mediated tumor multidrug resistance in rive with little influence on cytoehrome P-450 activities of rat liver.
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Three pSIREN-siRNA plasmids were constructed using a pSIREN-RetroQ vector to silence the expression of muhidrug resistance-associated protein (MRP1) gene, and subsequently characterized by restriction endonuclease digestion and DNA sequencing. A truncated MRP1 and a full-length MRP1 were cloned into pEGFP-N2 and PeDNA3.1 respectively as pEGFP-MRP1T and pcDNA-MRP1. The plasmid pEGFP-MRP1T was co-transferred with each of the three pSIREN-siRNAs into HEK293 cells for MRP1T-GFP targeted silencing, and pSIREN-siRNA1 was used as the negative control, pSIREN-siRNA2 and pSIREN-siRNA3 appeared to be more effective to silence MRP1T-GFP compared to pSIREN-siRNA1 as shown by fluorescence microscopy. For the silencing of full-length MRP1 expression, HEK293 ceils were co-transferred with pcDNA-MRP1 and either of the three pSIREN-siRNAs, then subjected for Western blot analysis and MTT assays, pSIREN-siRNA2 and pSIREN-siRNA3 were able to inhibit the expression of 190 kD MRP1, but not pSIREN-siRNA1. The MDR of MRP1-transfected HEK293 ceils was abolished with pSIREN-siRNA2 or pSIREN-siRNA3 transfections. RNA secondary structure predictions demonstrated that the mRNA local free energy (△G) of the siRNA1 targeted sequence was lower, as the GC content and Tm value of siRNA1 were higher than those of siRNA2 and siRNA3. These data suggest that the local structure siRNAs and target mRNA may influence the silencing efficiency of MRP1 expression.
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Objective To explore time difference attack therapy in Acinetobacter bauamnnii infection. Methods 67 patients with Acinetobacter bauamnnii were divided into two groups. The experimental group were first given Fosfomycin 4 g 5% GS 100 ml iv girt to finish within 60 min and then given Cefperazone/Sulbactam 4 g 0.9% NS 250 ml iv gitt immediately bid. The control group were given: Ampicillin/Sulbactam 3 g 0.9% NS 250 ml iv gitt (tid) + Ciprofloxacin 0.2 g iv girt (bid). The treatment course all was 11 days. Results The overall effective rate of experimental group methods was superior to that of control group(X2 =9.56 ,P =0. 023). Conclusion The Fos-fomycin Cefperazone/Sulbactam time difference attack therapy for the treatment of Acinetobacter the bauamnnii in-fection is a new way.
