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目的探讨~(131)I-17-丙烯胺基-17-去甲氧基格尔德霉素(17-AAG)对人乳腺癌细胞生长的影响及其相关机制。方法采用过氧化氢标记法制备~(131)I-17-AAG。细胞杀伤实验分为5组:二甲基亚砜(DMSO)对照(A)组,Na~(131)I 370 kBq(B)组,17-AAG 2.5 mg/L(C)组,~(131)I-17-AAG 370 kBq(D)组,~(131)I-17-AAG 370 kBq+17-AAG 2.5 mg/L(E)组。用四甲基偶氮唑蓝(MTT)法检测各种药物对人乳腺癌细胞 MCF-7的生长抑制作用,流式细胞术分析细胞凋亡及细胞周期变化,RT-PCR 检测药物处理前后 MCF-7细胞中 Akt2基因的 mRNA 表达情况。结果 ~(131)I-17-AAG 的标记率为83%,放化纯为96.6%,比活度为1.48×10~5MBq/μmol。各组药物对细胞的杀伤呈时间效应,随着时间的延长,细胞的抑制率都呈明显上升趋势,尤以 E 组趋势明显。A～E组药物作用48 h 后,通过亚 G1峰检测 MCF-7细胞凋亡率分别为(1.54±0.13)%,(5.72±1.05)%,(12.97±1.44)%,(20.65±1.36)%,(35.39±4.15)%,各组细胞凋亡率差异有统计学意义(P 均<0.05)。C 组,D 组及 E 组Akt2基因的 mRNA 表达均比 A 组降低,其中 E 组降低尤为明显。结论 ~(131)I-17AAG 能够抑制 MCF-7细胞的生长并促进其凋亡,且能有效抑制 Akt2基因的 mRNA 表达,和17-AAG 联合应用能够增强肿瘤细胞对放疗的敏感性。
Objective To investigate the effect of (131) I-17-acrylamido-17-demethoxygeldanamycin (17-AAG) on the growth of human breast cancer cells and its related mechanism. Methods ~ (131) I-17-AAG was prepared by hydrogen peroxide labeling. The cell killing assay was divided into five groups: DMSO control group (A), Na ~ (131) I 370 kBq (B) group, 17-AAG 2.5 mg / ) I-17-AAG 370 kBq (D), 131 I-17-AAG 370 kBq + 17-AAG 2.5 mg / L (E). The inhibitory effects of various drugs on the growth of human breast cancer cells MCF-7 were detected by MTT assay. The apoptosis and cell cycle were analyzed by flow cytometry. The levels of MCF -7 cells Akt2 gene mRNA expression. Results The labeling rate of ~ (131) I-17-AAG was 83%, the radiochemical purity was 96.6% and the specific activity was 1.48 × 10 ~ 5MBq / μmol. Each group of drugs on the cell killing was a time effect, with the extension of time, cell inhibition rates showed a clear upward trend, especially in the E group trend. The apoptosis rates of MCF-7 cells detected by sub-G1 peak were (1.54 ± 0.13)%, (5.72 ± 1.05)%, (12.97 ± 1.44)%, (20.65 ± 1.36) %, (35.39 ± 4.15)%, respectively. There was significant difference in apoptosis rate between groups (P <0.05). The mRNA expression of Akt2 in group C, group D and group E was lower than that in group A, especially in group E. Conclusions ~ (131) I-17AAG can inhibit the growth and promote the apoptosis of MCF-7 cells, and can effectively inhibit the expression of Akt2 mRNA. The combination of 17-AAG and 131-IAG can enhance the sensitivity of tumor cells to radiotherapy.