人卵巢癌COCl／5-Fu耐药细胞株多药耐药与凋亡关系的研究

The relative of multi-drug resistance and apoptosis of COC1/5-Fu

目的探讨耐药株人卵巢癌细胞COC1／5-Fu的耐药机制及耐药与细胞凋亡的关系。方法检测COC1／5-Fu细胞株对不同抗肿瘤药物的敏感性，流式细胞仪分析COC1／5-Fu细胞在5-Fu作用的凋亡情况以及对5-Fu的耐受分析，RT—PCR分析COC1／5-Fu细胞株中凋亡相关基因p53、bcl-2、bcl—xl、bax等基因的表达情况。结果COC1／5-Fu细胞与COCl细胞相比除对5-Fu耐药外，对其它几种常用化疗药均有不同程度的耐药性；COCl细胞未经5-Fu处理的对照组、30ixmol／L5-Fu组及150umol／L5-Fu组的凋亡指数与对照组相比，凋亡分别增加了1．2倍和2。1倍，差异有统计学意义（P〈0．05），而经同样浓度5-Fu处理的COC1／5-Fu细胞与其对照组相比，细胞凋亡变化不明显（P〉0．05）；经30umol／L和150umol／L5-Fu处理后，COC1细胞与其对照组相比，呈现明显的GO／G1期增加，S期、G2／M减少，而COC1／5-Fu细胞与其对照组相比细胞周期未见改变；PCR分析发现与COC1细胞相比，COC1／5．Fu细胞耐药株bcl-xl、bcl-xs和bax的表达增高，而p53和cpp32的表达减少，特别是p53基因表达明显减少（P〈0．05）。结论wtp53基因的突变与卵巢癌细胞的细胞周期的改变、耐药性的产生相关，是卵巢癌细胞COC1／5-Fu产生多药耐药性的机制之-。

Objective To explore the mechanism of multi-medicine drug resistance in human ovarian cancer cell line COC1/5-Fu. Methods The apoptosis and the tolerance of COC1/5-Fu cell induced by 5-Fu were analyzed by FACS. The expression of apoptosis related genes, such as p53, bcl-2, bcl-xl and bax, in COC1/5-Fu cell line were analyzed by RT-PCR. Results The COC1/5-Fu cell has some degree of drug resistance to 5-Fu and several other commonly used kind of chemotherapy medicine, among of which, drug resistance of 5-Fu reach 107.0 times and Paclitaxel reach 9. 0 times compared with COC1. When COC1 was treated with the concentration of 5-Fu （0umo/L, 30umo/L or 150 umol/L） , the A1 was （6. 5 ± 1.0）% ,（14. 0 ±4. 0）% and （20. 0 ±5.0）% , respectively. The rate of apoptosis increased 1.2 time and 2. 1 time, compared with not treated with 5-Fu, which were significantly different （ P 〈 0. 05 ）. But when COC1/5-Fu was treated with the same concentration of 5-Fu （ 30 umo/L or 150 umol/L）, the AI was （ 6.7 ± 0. 7 ） %, （7.1 ± 2. 2 ） % and （6. 5 ± 2.0 ） %. When treated with the same concentration of 5-Fu （30 umo/L or 150 umol/L） , the proportion of apoptosis was significantly increased, GO/ G1 phase was increased, and S and G2/ M phases were reduced in COC1 cells, but the proportion of apoptosis and cell cycle was not changed in COC1/5-Fu cells. The expression of bcl-xl , bcl-xs and bax mRNA were significantly increased and the expression of p53 and cpp32 mRNA were significantly decreased in resistant COC1/5-Fu cells , compared with COC1 cells. Conclusion wtp53 gene mutation is related with cell cycle change of ovary cancer cell and drug resistance, which is one of multi- medicine drug resistance mechanisms of COC1/ 5-Fu.