人脑星形细胞瘤原代细胞体外化疗药物敏感性实验研究

目的应用偶氮噻唑蓝比色法(MTT),检测常用化疗药物卡莫司丁(BCNU)、替尼泊甙(VM26)、多柔比星(ADM)、顺铂(CDDP)、环磷酰胺(CTX)单用及联合应用对人脑星形细胞瘤(AT)细胞的敏感性,比较不同类型(高分级,低分级,原发,复发)星形细胞瘤细胞对化疗药物敏感性的差别,以期对AT术后化疗提供合理、个性化的用药依据。方法 44例不同类型人脑星形细胞瘤细胞(术后病理证实为AT),在CO2培养体系中进行体外原代培养,差速贴壁法将其分离纯化,应用免疫荧光技术检测脑胶质纤维酸性蛋白(GFAP)对其鉴定;在分离纯化鉴定后的AT细胞中,加入不同浓度的化疗药物进行干预,MTT法测光密度值(OD值),计算敏感率。结果 38例AT标本成功进行了体外培养,培养成功率86%,高分级AT培养成功率最高;五种药物对AT细胞的抑制范围差异性很大,敏感性均不同;不同作用机制的药物联合应用对AT的敏感率高于其单一药物,单用与联合间有显著性差异(P<0.05);高分级及复发AT细胞对单一药物的敏感率均高于低分级及原发AT细胞,高分级与低分级间、原发与复发间差异有统计学意义(P<0.05),而联合用药时,高分级与低分级间、原发与复发间均无差异性(P>0.05)。结论 AT细胞可在体外成功地培养、纯化及鉴定,培养成功率与肿瘤病理类型密切相关;不同肿瘤个体对药物敏感性差异很大,联合用药可提高药物的敏感性,化疗药物对高分级及复发AT的敏感性比对低分级及原发的高;临床上应根据药敏结果,选择敏感性药物,行个体化化疗,以提高疗效,最大限度地减少副反应及肿瘤复发。

Experimental Study on Chemotherapeutic Sensitivity of Human Astrocytoma Cells in Vitro

Objective To detect the sensitivity of usual chemotherapeutic drugs （BCNU, VM26, ADM, CDDP, CTX） used alone or combination to astrocytoma （AT） cells with MTT assay and compare the drug sensitivity to different types of AT cells （HGA, LGA, primary, recurrent）, expecting to provide a reasonable basis for personalized post-operative chemotherapy. Methods 44 different types of AT samples （pathologically confirmed AT） were cultured in CO2 training system, isolated and purified with differential-adhesion method and identified by immunofluorescence assay in detecting of glial fibrillary acidic protein （GFAP）, then added different concentration chemotherapy drugs, tested optical density （OD value） with MTT assay and calculated their sensitivity rates. Results 38 AT samples were successfully cultured, and the success rate was 86%, which was the highest in HGA cells. The sensitivity rates of chemotherapy drugs of AT samples were different and the inhibition range was greatly varying. The combination of two therapy agents, with different mechanisms or different cell-cycle targets, showed more sensitivity than single one （P 〈0.05）. The sensitivity rates of drugs in the high-grade and recurrence group were higher than those in the low-grade and primary group （P 〈0.05）, but there were no statistical significance in combination chemotherapy to high-grade and low-grade group or primary and recurrent group （P 〉0.05）. Conclusions The success rate of AT cells culture is closely related to pathological types of tumor. The sensitivity of chemotherapeutic drugs to AT cells varies widely, even the same type of tumor （like pathological grade） has different sensitivity rates to the same drug. Combined chemotherapy can increase the drug sensitivity rate. Antineoplastic drugs have high sensitivity to high-grade and recurrent AT. Sensitive antineoplastic agents of clinical individualized chemotherapy may be selected according to the results of drugs test so as to increase treatment effect, minimize side effects and tumor recurrence.