miR-181d在调节小细胞肺癌化疗敏感性中的作用

Role of miR- 181d in the regulation of chemotherapy sensitivity in small cell lung cancer

目的观察miR-181d在调节小细胞肺癌(SCLC)化疗敏感性中的作用。方法选取2010年3月至2015年3月期间接受手术切除的SCLC患者的肿瘤组织标本112例作为研究对象,利用QRT-PCR以及Western blot法检测SCLC H69及H69AR内miR-181d的表达情况,以及上调H69AR内miR-181d的表达,测定miR-181d与BCL2的表达,利用CCK8测定细胞对于化疗药物产生的敏感性。结果 miR-181d在H69AR内的表达水平显著低于H69,且在化疗耐药性SCLC组织内的表达水平显著低于化疗敏感性SCLC组织,差异均有统计学意义(均P<0.05)。BCL2基因及蛋白在H69AR内的表达水平均显著高于H69,差异均有统计学意义(均P<0.05)。miR-181d在H29AR-e GFP-miR181d内的表达上调约7倍,且显著高于在H29AR及H29AR-e GFP-NC内的表达,差异有统计学意义(P<0.05)。BCL2基因及蛋白在H29AR-eG FP-miR181d内的表达均分别显著低于在H29AR及H29AR-eG FP-NC内的表达,差异有统计学意义(P<0.05)。H69AR转染eG FP-miR181d之后,对于顺铂和阿霉素以及依托泊苷产生的敏感性显著增加,且IC50值下降,差异有统计学意义(P<0.05)。H69转染miR-181d有关抑制剂之后,miR-181d在H69-miR181d-inhibitor的表达显著低于在H69及H69-inhibitor-NC中的表达,差异均有统计学意义(均P<0.05)。同时,H69-miR181d-inhibitor细胞对于化疗药物产生的敏感性显著降低,且IC50值增大,差异有统计学意义(P<0.05)。结论 MiR-181d在SCLC的化疗敏感性方面发挥重要作用,且有望成为评价SCLC疗效的一个指标。

Objective To observe role of miR- 181 d in the regulation of chemotherapy sensitivity in small cell lung cancer（ SCLC）.Methods 112 SCLC patients who underwent surgical resection in our hospital from March 2010 to March 2015 were selected as the research object. miR- 181 d expression in H69 and H69 AR cell lines were detected by quantitative reverse transcriptase- polymerase chain reaction（ QRT-PCR） and Western blot. After the up- regulation of miR- 181 d expression in H69 AR,the expressions of miR- 181 d and BCL2 was detected.The sensitivity of cells to chemotherapeutic drugs was determined by CCK8 assay. Results miR- 181 d expression in H69 AR was significantly lower than that in H69,and miR- 181 d expression in tissue specimens obtained from chemoresistant SCLC patients was significantly lower than those from chemosensitive SCLC patients（ all P〈0. 05）. The expression levels of BCL2 gene and protein in H69 AR were significantly higher than those in H69（ all P〈0. 05）. miR- 181 d expression in H29AR- e GFP- miR181 d was up- regulated about 7 times and it was higher than that in H29 AR and H29AR- e GFP- NC（ P〈0. 05）. The expressions of BCL2 gene and protein in H29AR- e GFP- miR181 d were significantly lower than those in H29 AR and H29AR- e GFP- NC（ P〈0. 05）. h69AR- e GFP- miR181 d significantly increased sensitivity to cisplatin,doxorubicin and etoposide,and IC50 values decreased（ P〈0. 05）. miR- 181 d expression in H69- miR181d- inhibitor was significantly lower than that in H69 and H69- inhibitor- NC（ all P〈0. 05）. At the same time,the sensitivity of H69- miR181d- inhibitor cells to chemotherapeutic drugs was significantly lower,and the IC50 values were increased（ P〈0. 05）. Conclusion MiR- 181 d plays an important role in the chemotherapy sensitivity of SCLC,and is expected to be an indicator for the treatment efficacy of SCLC patients.