洛铂对宫颈癌SiHa细胞射线敏感性的影响

Effect of lobaplatin on radiosensitivity of human cervical carcinoma SiHa cells

目的研究洛铂对宫颈癌SiHa细胞射线敏感性的影响及其机制。方法 MTT法检测不同浓度洛铂对SiHa细胞的作用24,48和72 h后的增殖情况,计算IC_(50)值。单克隆形成实验检测洛铂对SiHa细胞的射线敏感性影响,单击多靶模型拟合SiHa细胞的放射生物学参数。基于MTT法和细胞克隆形成实验结果,将细胞分为空白组(不加任何药物)和洛铂组(5.0μmol·L^(-1))、放疗组(6 Gy 6 MV X线)和洛铂+放疗组(5.0μmol·L^(-1)洛泊+6 Gy 6 MV X线)。流式细胞术检测洛铂、射线以及洛铂联合射线对SiHa细胞凋亡率的影响。Western blot检测促凋亡蛋白Bax和抑凋亡蛋白Bcl-2在SiHa细胞中的表达。结果空白组、洛铂组、放疗组和洛铂联合放疗组的凋亡率分别为(4.44±2.07)%,(30.50±3.17)%,(43.08±5.93)%和(84.68±4.90)%。洛铂组、放疗组相对空白组促凋亡蛋白Bax表达显著上调,抑凋亡蛋白Bcl-2显著下调。洛铂联合放疗组相对洛铂组、放疗组进一步显著上调Bax和下调Bcl-2的表达。结论洛铂可能通过细胞凋亡途径增强SiHa细胞射线敏感性。

Objective To investigate the effect of lobaplatin on the radiosensitivity of cervical cancer SiHa cells and its mechanism. Methods MTT assay was used to detect the proliferation of SiHa cells treated with different concentrations of lobaplatin at 24,48 and 72 h,and the IC_（50） values were calculated. The radiosensitivity of SiHa cells were measured by colony formation assay,and the multi-target single hit model was used to fit the radiological parameters of SiHa cells. Flow cytometry was used to detect the effect of lobaplatin,radiation and lobaplatin combined with radiation on the apoptotic rate of SiHa cells. Western blot was used to detect the expression of pro-apoptotic protein Bax and anti-apoptotic protein Bcl-2 in SiHa cells. Results According to the results of MTT assay and colony formation assay,cells were divided into four groups as follows： blank group（ routine cell culture）, lobaplatin group（ 5. 0 μmol·L^（-1）lobaplatin）,radiation group（ 6 Gy 6 MV X-ray） and lobaplatin ＋ radiation group（ 5. 0 μmol·L^（-1）lobaplatin ＋ 6 Gy 6 MV X-ray）. The apoptosis rates of the blank group,the lobaplatin group,the radiotherapy group and the combination group were（ 4. 44 ± 2. 07） %,（ 30. 50 ± 3. 17） %,（ 43. 08 ± 5. 93） % and（ 84. 68 ± 4. 90） %,respectively. The expression of pro-apoptotic protein Bax was significantly up-regulated and the expression of Bcl-2 was down-regulated in the lobaplatin group and the radiotherapy group compared with theblank group. Compared with the lobaplatin group and the radiotherapy group,the expression of pro-apoptotic protein Bax was significantly up-regulated while the expression of Bcl-2 was down-regulated in the combination group.Conclusion Lobaplatin might enhance the radiosensitivity of SiHa cells through the apoptosis pathway.