心肌细胞裂解液对鼻咽癌CNE-2细胞株的放射增敏作用及其机制的研究

摘要

Objective To study the effect of proliferation and radiation sensitivity of cardiac muscle cell lysate on human nasopharyngeal carcinoma CNE-2 cells. Methods Lysates of rat cardiac muscle tissue of > lOkD, <5kD and 5-10kD were obtained by ultrafsltration centrifugation method, including CMCLnr from neonatal Wisiar rat and CMCl.ar from adult Wistar rat; culture solutions of rat cardiac muscle tissue of > lOkD, <5kD and 5-10kD were obtained too, CMCMnr from neonatal Wistar rat and CMCM20dr from 20-day old neonatal Wistar rat. Survival rate of CNE-2 cells was detected when it was treated with 0. 3mg/ml different molecular weights of CMCLnr, CMCLar, CMCMnr, CMCM20dr and 1.26u.g/ml DDP for 48h. The radiosensibility of CMCLnr(5-)0kD) and topotecan (TPT) was detected and sing]e-hil multitarget model was used to plot survival curve. CNE-2 cell cycle and apoplosis were measured by flow cytometer. Results Compared with control group, 5-10kD CMCLnr, CMCLar, CMCMnr, CMCM20dr couid significantly inhibit the survival rate of CNE-2 cells (P < 0. 05 ) . They had the same anticancer activity as platinum (P > 0. 05 ) . The values of D0, Dt, N and SF2 of CNE-2 cells received radiation treatment alone were 1.185Cy, 0.676Cy, 3.729 and 69%. Those of CNE-2 cells received radiation plus CMCLnr (5-10kD) were 0. 762Cy, 0.600Gy, 6. 147 and 25%. SERPO , SERDq and SERSF2 were 1.555, 1. 127 and 2. 760. The apoptotic rate of CMCLnr + radiation group was ( 19. 34 ± 0. 23) % , which was higher than that in topotecan + radiation group and single radiation group( P < 0. 05). Conclusion The cardiac muscle cell lysate can inhibit the proliferation of CNE-2 cells and has shown radiosensitizing effect in vitro on CNE-2 cells. The radiosensitizing mechanism may be related to the induction of S phase arrest and apuptosis-promoling effects.%目的 探讨心肌细胞裂解液对鼻咽癌CNE-2细胞的放射增敏作用和机制.方法 采用超滤离心法获得分子量＞ 10kD、＜5kD及5～10kD的乳Wistar大鼠心肌细胞裂解液超滤液(CMCLnr)、成年Wistar大鼠心肌细胞裂解液超滤液( CMCLar)、乳Wistar大鼠心肌细胞培养液超滤液(CMCMnr)和20日龄乳Wistar大鼠心肌细胞培养液超滤液(CMCM20dr).用抑瘤活性实验检测上述滤液(0.3mg/ml)和顺铂(1.26 μg/ml DDP)作用鼻咽癌CNE-2细胞株的细胞存活率.用放射增敏实验检测CMCLnr(5～10kD)和拓扑替康(TPT)的放射(RT)增敏效果,单击多靶数学模型进行曲线拟合作图.流式细胞术检测TPT、CMCLnr(5～10kD)、RT单独或联合作用CNE-2细胞后细胞周期分布及凋亡率情况.结果 分子量为5～10kD CMCLnr、CMCLar、CMCMnr和CMCM20dr均较对照组能够显著降低CNE-2细胞的存活率(P＜0.05),且与DDP组差异无统计学意义.CNE-2细胞RT组的D0值为1.185Gy,Dq值为0.676Gy,N值为3.729,SF2为69％;CMCLnr+ RT组的D0值为0.762Gy,Dq值为0.600Gy,N值为6.147,SF2为25％,放射增敏比指标SERD0为1.555,SERDq为1.127,SERsP为2.760.CMCLnr+ RT组的细胞凋亡率为(19.34±0.23)％,均高于TPT+RT组和RT组,差异有统计学意义(P＜0.05).结论 心肌细胞裂解液可抑制鼻咽癌CNE-2细胞的增殖,并对CNE-2细胞具有较强的放射增敏作用,细胞水平的研究表明其放射增敏的机制可能与诱导S期阻滞及促进凋亡有关.