RT-PCR检测乳腺癌多药耐药基因表达及体外耐药逆转的研究

Quantitative analysis of multion resistance (mdr) gene expression in breast cancers by reverse transcription polymerase chain reaction (rt-pcr) and the reversal effects by therapeutic agents in vitro

目的 研究乳腺癌组织中多药耐药基因（ Ｍ Ｐ Ｒ１） 的表达及临床意义；探讨３ 种耐药逆转剂维拉帕来（ Ｖ Ｐ Ｌ） 、三苯氧胺（ Ｔ Ａ Ｍ） 、潘生丁（ Ｄ Ｐ） 对耐阿霉素人乳腺癌细胞系 Ｍ Ｃ Ｆ７ａｄｒ 的逆转作用。方法 采用逆转录—多聚酶链式反应技术（ Ｒ Ｔ － Ｐ Ｃ Ｒ）检测２６ 例乳腺癌病人 Ｍ Ｄ Ｒ１ 的表达情况；采用 Ｍ Ｔ Ｔ 比色法测定每孔细胞的吸光度 Ａ 值， 并计算细胞抑制率（ Ｉ Ｒ ％ ） 、半数细胞抑制浓度（ Ｉ Ｃ５０） 。结 果 乳 腺 癌 Ｍ Ｄ Ｒ１ 基 因 阳 性 表 达 率 为 ４６２ ％ （１２／２６） ；其中在未经任何治疗的１８ 例原发乳癌中 Ｍ Ｄ Ｒ１阳性率为２７８ ％ （５／１８） ；耐药基因表达与乳腺癌的组织学类型无相关性，但在有转移和有化疗史的乳癌中， 其阳性率要明显高于 无转 移和 无 化疗 史 者（ ７２７ ％ 、８７５ ％ 、２６７ ％ 、２７７ ％ ） 。 Ｍ Ｔ Ｔ 法证实，在无细胞毒性的剂量范围内， 分别合用 Ｖ Ｐ Ｌ、 Ｔ Ａ Ｍ和 Ｄ Ｐ 前后， Ｍ Ｃ Ｆ７ａｄｒ 细胞对阿霉素的敏感性有明显差异， Ｉ Ｃ５０ 降至原来的９ ％ ～５０ ％ 。结论 乳腺癌有一定的原发耐药性， 化疗能够促进 Ｍ Ｄ Ｒ１ 的表达，

Objective\ To study the expression MDR 1 gene in breast cancer and the reversal of MDR by Three therapelitic agents in vitro.Methods Surgical specimens from 26 cases of breast cancer were studied by RT-PCR to analyse the expression of MDR 1 gene.MTT assay were performed to measure the cytotoxity to MCF 7 adr cells by the combination of adriamycin (ADM) and one of the reversal agent (verapamil, tamoxifen and dipridamole) respectively.Results\ The total MDR1 gene positive rate were 46.2% (12/26).it was much higer than those without any treatment (26.6% 5/18) The positive MDR 1 gene expression were not related with their histolgical pattern of the tumors (P>0 05).This study showed MDR 1 were higher in the metastasis. patients(72.7%∶27.7%,P<0.05),or chemotherapy group (87.5%:27.7%,P<0.05).It was found above three reversal agents could enchance the sensitivity of MDR 1 cells to ADM.Conclusion There exists nature drug resistance in breast cancer to some extent. Gene expression can be increased by chemotherapy or metastasis All three therapeutic agents in this study can reverse the multidrug resistomce of MCF 7 adr cells in vitro.