Objective To compare the positive rate of zinc finger protein A20， NF-κB p65 protein， and P-glyco- protein between primary hepatocellular carcinoma （HCC） tissues and paratumor tissues， and to explore the relationship between the 3 kinds of proteins and pathological features of HCC. Methods Thirty-two HCC tissues and 26 paratumor tissues resected from patients with HCC treated in our hospital from Feb. 2009 to Aug. 2010 were enrolled. Clinical data were also collected from files. The expressions of zinc finger protein A20， NF-κB p65 protein， and P-glycoprotein were tested by immunohistochemistry. Results The positive rate of zinc finger protein A20， NF-κB p65 protein， and P-glycoprotein in HCC tissues were 87.5%（28/32）， 81.3%（26/32）， and 65.6%（21/32）， respectively， which were higher than that in paratumor tissues〔61.5%（16/26）， 34.6%（9/26）， and 30.8%（8/26）， respectively〕， P＜0.05. The three kinds of proteins were all closely related with HbsAg， and zinc finger protein A20 was related with cirrhosis in addition （P＜0.05）. Conclusions The positive rate of zinc protein A20， NF-κB p65 protein， and P-glycoprotein are much higher in primary HCC tissues than that in paratumor tissus， and they may play an important role in preoperative determination of hepatic tumors.
【Abstract】ObjectiveTo construct an mdr1 expression vector and detect its expression in HepG2 cells in vitro. MethodsThe 4.5-kb mdr1 cDNA was obtained from the plasmid pHaMDR1 cloned into the PCIneo mammalian expression vector, which was later transferred into human hepatocarcinoma cell line HepG2 by liposome. Then the HepG2 cells resisting G418 were clustered and proliferated，and the specific fragment of mdr1 cDNA, mRNA and the Pgp in these HepG2 cells were detected by means of PCR, RT-PCR and FCM respectively. ResultsThe mdr1 expression vector was constructed successfully，and the stable multidrug resistance(MDR) hepatocarcinoma cell line （HepG2/mdr1） was developed as well. The outcome of PCR analysis showed that the specific fragment of mdr1 cDNA could be found in HepG2/mdr1 cells, but not in the nontransfection HepG2 cells. Furthermore,the content of the specific fragment of mdr1 mRNA and the expression of P-gp in HepG2/mdr1 cells were (59.7±7.9)% and (12.5±5.45)% respectively, the corresponding value in HepG2 cells were (16.9±3.2)% and (4.63±2.59)% respectively. The difference was statistically significant (P＜0.05). ConclusionIt is praticable to develop MDR hepatocarcinoma cell line by transferring mdr1 cDNA into HepG2 cells, which is useful in the research of MDR mechanism.
【Abstract】ObjectiveTo study the expressions of P-gp, nm23 and p53 in primary breast cancer tissues and to evaluate the prediction significance in recurrence of the primary breast cancer. MethodsExpressions of P-gp, nm23 and p53 in 57 benign and malignant breast paraffin-embedded specimens, the difference of the rate and intensity of positive reaction of P-gp,nm23 and p53 were checked by immunohistochemical staining.ResultsThe positive rate of nm23 significantly increased in benign breast tissues compared with the tissues of recurrence of breast cancer. The positive intensity of nm23 was significantly decreased in the tissues of recurrence of breast cancer compared with the tissues of primary breast cancer. The positive rate of p53 was significantly increased in malignant breast tissues compared with benign breast tissues. The positive intensity of p53 was significantly increased in the tissues of recurrence of breast cancer compared with the tissues of primary breast cancer. There was no difference in the positive rate of P-gp in the tissue of recurrence and primary of breast cancer compared with benign breast tumor. But the positive intensity of P-gp was significantly decreased in the tissues of recurrence and primary of breast cancer compared with benign breast tumor. ConclusionThe expression of p53 may indicate the proliferating activity of carcinoma cell. The positive intensity of expression of p53 and nm23 is valuable in prediction of the recurrence of primary breast cancer.
Multidrug resistance (MDR) remains the major obstacle to the success of clinical cancer chemotherapy. P-glycoprotein (P-gp), encoded by the MDR1, is an important part with complex mechanisms associated with the MDR. In order to overcome the MDR of tumors, we in the present experimental design incorporated small interfering RNA (siRNA) targeting MDR1 gene and anticancer drug paclitaxel (PTX) into the solid lipid nanoparticles (SLNs) to achieve the combinational therapeutic effects of genetherapy and chemotherapy. In this study, siRNA-PTX-SLNs were successfully prepared. The cytotoxicity of blank SLNs and siRNA-PTX-SLNs in MCF-7 cells and MCF-7/ADR cells were detected by MTT; and the uptake efficiency of PTX in MCF-7/ADR cells were detected via HPLC method; quantitative real-time PCR and flow cytometry were performed to investigate the silencing effect of siRNA-PTX-SLNs on MDR1 gene in MCF-7/ADR cells. The results showed that PTX loaded SLNs could significantly inhibit the growth of tumor cells, and more importantly, the MDR tumor cells treated with siRNA-PTX-SLNs showed the lowest viability. HPLC study showed that SLNs could enhance the cellular uptake for PTX. Meanwhile, siRNA delivered by SLNs significantly decreased the P-gp expression in MDR tumor cells, thus increased the cellular accumulation of rhodamine123 as a P-gp substrate. In conclusion, the MDR1 gene could be silenced by siRNA-PTX-SLNs, which could promote the growth inhibition efficiency of PTX on tumor cells, leading to synergetic effect on MDR tumor therapy.
Objectives The purpose of this study is to verify the phenytoin-resistant mesial temporal lobe epilepsy (MTLE) induced by Li-pilocarpine and screened by antiepilepsy drug (AEDs). Methods The rats with MTLE were induced by Li-pilocarpine, which were screened by effect of phenytoin treatment monitored by vedio-EEG. The living microdialysis technology was used for verification of drug concentration in brain of drug-resistant and drug-responsive rat model, and the P-glycoprotein expression was detected by immunohistochemical method. Results Sixteen rats with chronic MTLE were successfully induced in total 30 rats, among which, 6 drug-resistant rats with MTLE were screened. The brain/plasma ratio of area under the curve in drug-resistant rats was significantly lower than that of drug-responsive rats (0.15±0.03 vs. 0.28±0.05, P<0.05). In addition, the P-glycoprotein expression in brain of drug-responsive rats was obviously higher than that of drug-responsive rats (P<0.05). Conclusions The low concentration of phenytoin in drug-resistant rat model with MTLE was verified that might be related to the over-expressed P-glycoprotein in brain.
ObjectiveIn order to evaluate that whether the P-glycoprotein-inhibitor verapamil (VPM) could effect the distribution of antiepileptic drug phenytoin (PHT) in a rat model of mesial temporal lobe epilepsy (MTLE).MethodsThe rat models of MTLE were induced by li-pilocarpine and were randomly divided into two groups (PHT group and VPM+PHT treatment group) to compare the PHT distribution in brain, liver and kidney. Brain dialysate samples were collected by microdialysis technology. And the analysis of samples for PHT concentration was performed by high performance liquid chromatography (HPLC). The comparisons were carried out by t test (or Wilcoxon test).ResultsIn VPM+PHT treatment group, 4 out of 9 rats were dead within 30 minutes after drug administration. The significantly decreased area under the curve (AUC) ratio of brain/plasma in VPM+PHT group was 0.11±0.06 when compared with PHT group 0.21±0.02 (t=3.237, P=0.025), while there were no significant differences in ratios of liver/plasma ［PHT (1.12±0.37) vs. VPM+PHT (0.99±0.27), Z=−0.490, P=0.624］ and kidney/plasma ［PHT (0.74±0.16) vs. VPM+PHT (0.49±0.26), t=1.872, P=0.103］ between two groups.ConclusionsThe P-glycoprotein-inhibitor VPM significantly decreased PHT level in brain of rat with MTLE.