【Abstract】ObjectiveTo construct a mrp1 expression vector and investigate its biological characteristics in HepG2 cells in vitro. MethodsThe 6.5 kb multidrug resistanceassociated protein （MRP) cDNA obtained from plasmid pGEM-mrp1 was cloned into the pCI-neo 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 mrp1 mRNA and MRP in these HepG2 cells were detected by means of RT-PCR and FCM respectively. ResultsThe mrp1 expression vector was established successfully, and the stable MDR hepatocarcinoma cell line (HepG2/mrp1) was developed as well. The content of the specific fragment of mrp1 mRNA was (56.8±6.37)% and MRP was 7.89 in the HepG2/mrp1 cells, the corresponding value in HepG2 cells was (9.67±3.26)% and 0.79 respectively. The difference was statistically significant (P＜0.05). ConclusionIt is practicable to establish MDR hepatocarcinoma cell line by transferring mrp1 cDNA into HepG2 cells, which is useful in the research of MDR mechanism.
Objective To review the application advancements of ATP-binding cassette (ABC) transporter in medical research.Methods Relevant literatures about the applications of ABC families in medical research were reviewed. Results ABC families mainly took roles in transporting substances across cell membrane. Some of them were useful for the prediction of drug resistance and the prognosis of malignant tumors. Others were target s for molecular researches. Their expressions or mutations might be related with the occurrence of diseases. Conclusion ABC families are very important in the diagnosis and therapy for diseases. Thus they are very promising tools for future medical research.
ObjectiveTo summarize the latest advances in copper and cuproptosis in the field of breast cancer, and to provide a reference for clinical treatment decisions. MethodThe literatures related to copper and cuproptosis in recent years were read and summarized, and the research progress on the role of copper in breast cancer, the application of cuproptosis in the diagnosis and treatment of breast cancer were reviewed. ResultsCuproptosiswas a novel form of programmed cell death, which occurred via direct binding of copper to lipoylated components of the tricarboxylic acid (TCA) cycle, this resulted in lipoylated protein aggregation and subsequent iron-sulfur cluster protein loss, leading to proteotoxic stress and ultimately cell death. Cuproptosis induced proliferation and migration of breast cancer cell , mediated personalized immunotherapy, and participated in endocrine and chemotherapeutic drug resistance. ConclusionExploring the mechanism of cuproptosis provides potential applications for subsequent immunotherapy, endocrine therapy, and chemotherapy for breast cancer, leading to new effective strategies for patients.
Objective To dynamically study the formation of multidrug resistance(MDR) of human hepatocellular carcinoma cell SMMC-7721 induced by Adriamycin (ADM) and the role of multidrug resistance-associated protein(MRP) in its mechanisms.Methods Hepatocellular carcinoma cell SMMC-7721 was cultured in RPMI-1640 medium containing ADM with progressively increased concentration or directly cultured in medium containing different concentrations of ADM. Resistant index of drug-resistant variants of SMMC-7721 cell was determined by drawing cell dosage-reaction curves.Levels of MRP mRNA expression were detected by reverse transcription-polymerase chain reaction(RTPCR). Intracellular rubidomycin(DNR) concentration was examined by flow cytometry(FCM).Results With progressive increasing of ADM concentration in medium resistant index and levels of MRP mRNA expression were correspondingly increased but intracellular DNR concentration was markly reduced. When parental cells were directly cultured in medium containing different concentrations of ADM, the higher the ADM concentration, the higher the level of MRP mRNA expression, but intracellular DNR concentration was kept at the similar high level and most cells died. Conclusion ADM may progressively induce SMMC-7721 cell resistant to multiple chemotherapeutic drugs with reduced intracellular DNR accumulation associated with the overexpression of MRP gene.
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.
This paper aims to study the effects of traditional Chinese medicine Euphorbia esula on multidrug resistant human gastric cancer cells in the cell proliferation, migration, invasion and apoptosis, and to study the apoptosis-inducing pathway. Different dilutions of Euphorbia esula extract were used to process human multidrug resistant gastric cancer SGC7901/ADR cells. Cell proliferation inhibition phenomenon was determined by MTT experiment. Nuclear morphological changes of apoptotic cells and apoptotic indexes were observed and determined by Hochest33528 staining followed with fluorescence microscope observing. Flow cytometry was used to detect cell apoptosis rate. Cell migration and invasion ability were observed and determined by Transwell method. Spectrophotometry was used to detect caspase-3 and caspase-9 enzyme activity. Western blotting was used to detect subcellular distribution of cytochrome c. The results showed that Euphorbia esula extract had obvious inhibition effect on proliferation of gastric cancer multidrug resistant SGC7901/ADR cells, which was time- and concentration-dependent. After processing multidrug resistant gastric cancer SGC7901/ADR cells with Euphorbia esula extract, the apoptotic index and apoptosis rate were significantly increased than those in the control group, which showed a time- and dose-dependent mode; but if a caspase inhibitor was added, apoptosis index was not obviously increased. Transwell method showed that migration and invasion ability of the Euphorbia esula extract-processed SGC7901/ADR cells dropped significantly. Spectrophotometry showed that in Euphorbia esula extract-processed SGC7901/ADR cells, caspase-3 and caspase-9 expression were increased, which had significant differences with the control group. Western blotting test showed that the distribution of cytochrome c decreased in mitochondria, while increased in the cytoplasm (i.e., cytochrome c escaped from mitochondria to the cytoplasm). In conclusion, Euphorbia esula extract could inhibit the proliferation, migration and invasion, and induce apoptosis in human gastric cancer multidrug resistant SGC7901/ADR cells; and cytochrome c, caspase-9 and caspase-3 might be involved in cell apoptosis induced by Euphorbia esula extract, suggesting endogenous or mitochondrial apoptotic pathway.
Objective To investigate the reversal effect of antisense phosphorothioate oligonucleotide （ASOND) on human hepatoma resistant cells. Methods Human hepatoma resistant cells SMMC-7721 was transfected with synthetic antisense phosphorothioate oligonucleotide complementary to the 5′ region flanking the AUG initiation codon mediated by lipofectamine. In vitro drug sensitivity was measured by MTT assay. The expression of P-170 was determined by flow cytometry and mRNA was assessed by RT-PCR. Results ASOND inhibited the expression of mRNA and p-170 in SMMC-7721, enhanced the sensitivity of SMMC-7721 to chemotherapeutic drug. The best inhibitory effect was achived by the dose of 0.5μmol/L. Conclusion ASOND enhanced the sensitivity of SMMC-7721 to chemotherapeutic drug and reversed the multidrug resistance of SMMC-7721 partially.
ObjectiveTo learn the status quo and characteristics of multi-drug resistant organism (MDRO) infection in a comprehensive hospital of the first grade in Sichuan Province, analyze the effect of prevention and control intervention, in order to provide a scientific basis for clinical MDRO prevention and control. MethodsWe collected MDRO data from January to June 2014 and from January to June 2015 through multi-drug resistance reporting software, and analyzed and compared the infection of MDRO during those two time periods. Then, we evaluated the prevention and control effect of MDRO infection. ResultsThe number of inpatients from January to June 2014 was 24709, among which 813 were detected with MDRO infection. Of those infected patients, 196 had nosocomial infection of MDRO and the other 617 had community infection/colonization. The proportion of nosocomial MDRO infection was 24.10%. The MDRO nosocomial infection case rate was 0.79%. The proportion of community MDRO infection/colonization was 75.90%. The number of inpatients from January to June 2015 was 25329, and 739 of them were found with MDRO infection, of whom 132 had nosocomial infection and 607 community infection/colonization. The proportion of nosocomial MDRO infection was 17.86%. The MDRO nosocomial infection case rate was 0.52%. The proportion of community infection/colonization was 80.14%. Compared with the first half of 2014, the proportion of nosocomial MDRO infection was lower with a statistically significant difference (χ2=9.062, P<0.001), and MDRO nosocomial infection case rate was also significantly lowered (χ2=14.220, P<0.001). There were significant differences between the first half of 2015 and the same period of 2014 in hospital department distribution of MDRO infection, patient infection site distribution and pathogen detection. ConclusionThe nosocomial MDRO infection control situation of our hospital is improved after the comprehensive prevention and control interventions, and we should focus on the prevention and control of key departments, important infection sites and major resistant bacteria in the future MDRO hospital infection control work.
Objective To investigate the clinical characteristics and drug resistance changes of nosocomial infection caused by carbapenem-resistant Klebsiella pneumoniae (CRKP) in different types of clinical departments, and to provide evidence for prevention and control of CRKP infection. Methods The hospital infection real-time monitoring system was used to retrospectively collect the inpatients with CRKP nosocomial infection in the First People’s Hospital of Lianyungang from January 2019 to December 2023 as the research objects. According to the different sources of departments, they were divided into intensive care unit (ICU) group, internal medicine group and surgery group. The changes of clinical characteristics and drug resistance to common antibiotics were analyzed. Results A total of 636188 inpatients were monitored, and 225 cases were infected with CRKP, with an overall infection detection rate of 0.035%. The detection rates of CRKP infection in the ICU group, internal medicine group, and surgery group were 0.736% (138/18749), 0.013% (44/336777), and 0.015% (43/280662), respectively, with the ICU group demonstrating a significantly higher rate than the other groups (P<0.05). The detection rates fluctuated in the early stage and then decreased rapidly in different years. The main infection site of CRKP in all groups was lower respiratory tract, but the proportion of device-related infections in the ICU group was higher than that in the internal medicine and surgery groups (P<0.05). In terms of the infected population, there was no significant difference in gender among groups (P>0.05) with the proportion of males more than 60%, while the difference in the proportion of patients aged ≥65 years among groups was statistically significant (P<0.05), with the highest in the internal medicine group (86.36%). The burden of underlying diseases and invasive operation exposure of the infected patients were high, and the proportion of cardiovascular and cerebrovascular diseases and indwelling catheters were as high as 69.33% and 83.56%, respectively. The differences in the proportions of cardiovascular and cerebrovascular diseases, diabetes mellitus, ≥3 underlying diseases, and surgical and invasive procedures among groups were statistically significant (P<0.05). The distribution of infection specimens in each group showed no statistically significant difference (P>0.05), with sputum, blood, and mid-stream urine specimens being the main detected specimens in all groups. The resistance rates of CRKP to penicillins and cephalosporins were more than 93%, and the resistance rates to aminoglycosides and sulfonamides were relatively low and showed a decline year by year. The resistance rate to ceftazidime/avibactam was only 7.41%, but the resistance rate to tigecycline increased. The difference in resistance rate of CRKP to co-trimoxazole among groups was statistically significant (P<0.05), while the differences in resistance to other antimicrobial agents were not statistically significant (P>0.05). Conclusions The detection rate, clinical characteristics and drug resistance of CRKP infection in different types of departments of medical institutions are different and changing. It is necessary to strengthen the rational use of antibiotics and the prevention and control of nosocomial infection.
Objective To review the recent studies on the multidrug resistance of breast cancer. Methods The literatures of recent years on the studies of multidrug resistance, multidrug resistance protein and breast cancer resistance protein were reviewed. Results Multidrug resistance resulted from multiple factors. How to identify the sensibility of chemotherapy drugs and select individual therapeutic regime early were important to improve the survival rate and life quality of breast cancer patients. Conclusion These studies on multidrug resistance of breast cancer are helpful to predicting the effect and outcome of chemotherapy and overcoming the barrier of drug resistance.