急性白血病患者骨髓间隙连接蛋白43的表达及其与耐药的相关性研究

Research on drug resistance and expression of connexin 43 in bone marrow of patients with acute leukemia

目的探讨急性白血病（AL）患者骨髓细胞中间隙连接蛋白43（Cx43）、P-糖蛋白（P-gp）及环氧合酶2（COX-2）基因表达水平及其与AL病程、预后和耐药的关系。方法77例不同病期AL患者，其中初治36例，完全缓解20例，复发20例，同时以20例异体造血干细胞移植供体及非血液系统恶性病患者为对照。采用SYBR Green实时定量反转录聚合酶链反应（SYBR—RT—PCR）技术，检测骨髓单个核细胞中Cx43、P—gp、COX-2 mRNA的表达，并对37例初治患者进行动态随访。结果AL初治组Cx43、P-gp、COX-2 mRNA的表达分别为0．52±0．57、1．42±1．06、1．14±0．95，复发组分别为0．20±0．40、2．29±1．11、1．69±0．81，完全缓解组分别为0．95±0．37、0．93±0．73、0．79±0．58，对照组分别为1．16±0．67、0．86±0．63、0．61±0．57。初治、复发AL患者Cx43 mRNA表达水平较对照组及完全缓解组低，差异均有统计学意义（初治组分别P=0．001、0．005；复发组均P〈0．001）；完全缓解组Cx43 mRNA表水平与对照组相比，差异无统计学意义（P=0．185）。AL患者骨髓细胞液中Cx43 mRNA与P—gp和COX-2 mRNA表达呈负相关，且初治组、完全缓解组及复发组表达均呈负相关（与P—gpr值分别为-0．471、-0．362、-0．526；与COX-2r值分别为-0．479、-0．344、-0．471）。36例AL初治患者随访4个月，死亡8例，生存28例，死亡患者Cx43 mRNA表达低于生存患者，差异有统计学意义（t=2．16，P=0．042）。结论初治、复发难治AL患者骨髓中Cx43 mRNA的表达下调，同时多药耐药基因P-gp、COX-2mRNA的表达上调；Cx43过度表达是预后良好因素，Cx43与AL的疗效、预后及化疗耐药密切相关。

Objective To detect the expression levels of connexin43（Cx43）, P-gp and COX-2 in bone marrow of patients with acute leukemia （AL）, and investigate their relationship with the pathogenesis, prognosis and resistance of this condition. Methods Using SYBR green real-time quantitative reverse transcription polymerase chain reaction （SYBR-RT-PCR）,the expression of Cx43, P-gp and COX-2 mRNA were detected in 77 AL patients at different phases, including 36 initially-treated, 20 complete-remission （CR） and 20 relapsed. A follow-up was done in those initially treated. Results Expression levels of Cx43, P-gp and COX-2 mRNA from initially-treated were 0.52±0.57, 1.42 ±1.06, 1.14±0.95;those from relapse AL patients were 0.20±0.40, 2.29 ±1.11, 1.69±0.81, respectively, those from CR patients were 0.95±0.37, 0.93±0.73, 0.79±0.58,respectively,those from normal patients were 1.16 ±0.67, 0.86±0.63, 0.61±0.57. Expression levels of Cx43 mRNA in initially-treated and relapse AL patients were significantly lower than those in normal patients and CR patients （initially-treated P were 0.001, 0.005;relapse patients were P〈0.001）,while the comparison between normal patients and CR patients showed no statistical significance （P=0.185）. Cx43 mRNA expression level was negatively correlated with P-gp and COX-2 mRNA, and a negative correlation was noted of both two expressions in initially-treated and relapse groups （Cx43 mRNA and P-gp mRNA r were -0.471, -0.362, -0.526;Cx43 mRNA and COX-2 mRNA r were -0.479, -0.344, -0.471）.A follow-up of four months was conducted in 36 initially-treated patients, eight of them died, The expression Cx43 in those who died was lower than that who survived. The difference was significant（t=2.16, P=0.042）. Conclusion Cx43 mRNA expression levels of initially-treated and relapse AL patients were lower than those in normal patients and CR patients, P-gp and COX-2 mRNA expression levels of initially-treated and relapse AL patients were higher than those in normal patients and CR patients. Cx43 expression probably is a favorable prognostic factor.Cx43 is participation in the genesis, development and drug resistance of AL.