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本研究探讨急性淋巴细胞白血病患儿骨髓间充质干细胞(mesenchymal stem cells,MSC)对K562/A02细胞耐药性的影响及其机制。从白血病患儿骨髓中分离、培养并鉴定MSC;建立K562/A02细胞株与MSC共培养的体系,应用AnnexinV-FITC检测一定浓度的阿霉素(ADM)对不同培养条件下K562/A02细胞凋亡的影响;RT-PCR检测K562/A02细胞中的凋亡基因家族中bcl-2和bax基因;荧光定量PCR检测多药耐药基因mdr1浓度。结果表明,单独培养组白血病细胞早期凋亡率为(8.38±0.29)%,而黏附培养组为(1.97±0.11)%,差异具有统计学意义(p<0.05)。相对于单独悬浮培养的K562/A02细胞,黏附共培养的K562/A02细胞bcl-2基因表达明显增强,bcl-2/bax比值显著增高;K562/A02单独悬浮培养组、黏附共培养组细胞中mdr1的水平比较无统计学差异(p>0.05)。结论:白血病患儿MSC能够使白血病细胞逃避药物的促凋亡作用,K562/A02对阿霉素产生耐药性可能与黏附共培养后bcl-2基因表达增强有关,而与其本身含有的mdr1基因无关。
This study was aimed to investigate the effects of mesenchymal stem cells (MSCs) on the drug resistance of K562 / A02 cells in children with acute lymphoblastic leukemia and its mechanism. Apoptosis of K562 / A02 cells was detected by Annexin V-FITC assay, and the concentration of doxorubicin (ADM) The expression of bcl-2 and bax in apoptosis gene family in K562 / A02 cells was detected by RT-PCR. The mdr1 concentration in multidrug resistance gene was detected by real-time PCR. The results showed that the early apoptosis rate of leukemia cells in cultured group was (8.38 ± 0.29)%, while it was (1.97 ± 0.11)% in adhesion culture group, the difference was statistically significant (p <0.05). Compared with K562 / A02 cells cultured alone, the expression of bcl-2 gene in co-cultured K562 / A02 cells was significantly increased and the ratio of bcl-2 / bax was significantly increased. In K562 / A02 suspension alone group and in co-culture group mdr1 levels were no significant difference (p> 0.05). CONCLUSION: MSC in leukemia can induce leukemia cells to avoid drug-induced apoptosis. The drug resistance of K562 / A02 to doxorubicin may be related to the increase of bcl-2 gene expression after co-culture, but not with its own mdr1 gene Nothing to do