ABSTRACT
OBJECTIVE@#To investigate the effecr of siRNA-interfering β-catenin expression on drug-resistance of multiple myeloma cells.@*METHODS@#The multiple myeloma cell line RPMI-8226 was cultured in vitro. The maphalan-resistant cell model was established by concentration gradient ascending of durg, then the drug-resistant cell line was instantaneously transfected with β-catenin siRNA, the sensitivity of RPMI 8226 cells to maphalan was detected by CCK-8 meltod before and after the transfection with siRNA; the mRNA and protein expression of β-catenin was detected by qRT-PCR and Western blot respectively, the apoptosis of cells was detected by flow cytometry.@*RESULTS@#IC of maphalan decreased from (5.29±0.19) μmol/L to (1.88±0.64) μmol/L, suggesting that the deplation of β-eatenin restored the sensitivity of drug-resistant cell line RPMI-8226 to malphalan. The Western blot showed that after the instaintaneous transfection with β-catenin siRNA, the β-catenin protein expression level obviously decreased, compared with level before transfection. After transfection, the maplalan-inducing apoptosis rate of cells increased from (35±0.5)% to (54±0.4)%, suggesting that the β-catinin gene may correlated with drug-resistance of cells. Interfering the expression of β-catenin gene could enhance the sensitivity of drug-resistant RPMI-8226 cells to maphalan.@*CONCLUSION@#The β-catenin siRNA interfereuce can inhisit the β-catenin gene expression in Wnt/β-catenin signaling pathway, suppress the cell proliferation, enhence the toxicity of maphalan on drug-resistant RPMI-8226 cells, thus result in increase of cell apoptosis.
Subject(s)
Humans , Apoptosis , Cell Line, Tumor , Cell Proliferation , Multiple Myeloma , RNA, Small Interfering , beta Catenin , GeneticsABSTRACT
OBJECTIVE@#To explore the effect of Bushen Yanggu Decoction (BYD) on drug resistance and proliferation of human multiple myeloma-resistant KM3/BTZ cells.@*METHODS@#Human multidrug-resistant KM3/BTZ cells were established by Bortezomib (BTZ) gradient induction. The effects of commonly used chemotherapeutic drugs and serum containing Bushen Yanggu Decoction (BYD) on the proliferation of KM3 cells and KM3/BTZ cells were detected by MTT assay. RT-qPCR and Western blot were used to detect the expression of Par-4, HSP27 and P-gp genes. Flow cytometry was used to detect cell apoptosis.@*RESULTS@#The established KM3/BTZ cells could produce varying degree of resistance to commonly used chemotherapeutic drugs. Among them, the highest resistance index (RI) to BTZ was 20.269. MTT assay showed that the proliferation of KM3/BTZ cells treated with serum containing Bushen Yanggu Decoction was inhibited, and the inhibitory effect increased with the serum concentration incranse of Bushen Yanggu Decoction. The serum containing Bushen Yanggu Decoction could inhibit the proliferation of KM3/BTZ cells, and induce apoptosis, significantly reduce the drug-resistance of KM3/BTZ cells, up-regulate the expression of Par-4, down-regulate the expression of HSP27 and P-gp.@*CONCLUSION@#Bushen Yanggu Decoction can effectively inhibit the proliferation of KM3/BTZ cells and induce apoptosis. Bushen Yanggu Decoction can effectively reverse the multidrug-resistance of KM3/BTZ cells. The mechanism may be related with the decrease of expression of HSP27 and P-gp and the increase of expression of Par-4.
Subject(s)
Humans , Apoptosis , Bortezomib , Cell Line, Tumor , Drug Resistance, Multiple , Drug Resistance, Neoplasm , Multiple MyelomaABSTRACT
Objective To study the effects and mechanisms of BuShenYangGuTang on proliferation and apoptosis of multiple myeloma cell line KM3. Methods The inhibitory effect of BuShenYangGuTang on cell proliferation was assessed by CCK-8. BuShenYangGuTang induced KM3 cell cycle arrest was analyzed by flow cytometry after propidium iodide staining. Flow cytometry was also used to analyze the cell apoptosis after Annexin V-FITC staining. The expressions of Bcl-2, Bax and NF-κB were determined by RT-qPCR and Western blot assay. Results BuShenYangGuTang inhibited the KM3 cell proliferation in a dose-dependent manner. The expression levels of Bcl-2 and NF-κB were decreased, the expression level of Bax was increased, and the cell cycle was arrested in G0 / G1 phase after treatment with BuShenYangGuTang. Conclusion BuShenYangGuTang could inhibit the proliferation, arrest cell cycle and induce the apoptosis in KM3 cells, which may be related to the abnormal expressions of Bcl-2, Bax and NF-κB.
ABSTRACT
<p><b>OBJECTIVE</b>To investigate the reversing effect of icaritin on multidrug resistance of multiple myeloma cell lines KM3/BTZ and its underlying mechanism.</p><p><b>METHODS</b>KM3/BTZ cells were established by a gradually ascending gradient induction of bortezomib (BTZ). The sensitivities of KM3 and KM3/BTZ cells to 7 chemotherapeutic drugs, the inhibition and reversal effects of icaritin on proliferation and drug-resistance of KM3/BTZ cells were analyzed by MTT. The apoptosis was analyzed by flow cytometry, and the expression of Par-4, HSP27 and P-gp were detected by Western blot.</p><p><b>RESULTS</b>KM3/BTZ cells were not only resistant to BTZ, but also to other 6 chemotherapeutic drugs. The resistance index (RI) to BTZ was 17.84, and higher than that of other chemotherapeutic drugs. Icaritin inhibited the proliferation and induced the apoptosis of KM3/BTZ cells. The ICvalue of BTZ decreased from 0.345 µg/ml to 0.149 µg/ml, and the reversal index was 2.38 (P<0.05). The expression of Par-4 protein increased in a concentration-dependent manner, while the expression of HSP27 and P-gp were down-regulated.</p><p><b>CONCLUSION</b>Icaritin can inhibit cell proliferation and induce apoptosis of KM3/BTZ cells, moreover, can effectively reverse the multidrug resistance of KM3/BTZ cells. The mechanism may be related with down-regulation of HSP27 and P-gp expression, and up-regulation of Par-4 expression.</p>