绒癌细胞株BeWo合体化后对不同化疗药物敏感性变化的研究

Changes of sensitivity of choriocarcinoma cell line BeWo to different chemotherapy drugs after fusion

目的通过检测绒癌细胞株Bewo合体化前后对不同化疗药物敏感性的差异，探讨滋养细胞合体化与恶性滋养细胞肿瘤耐药的关系，为临床恶性滋养细胞肿瘤，尤其是耐药恶性滋养细胞肿瘤的治疗提供新的思路和方法。方法利用forsoklin诱导绒癌细胞株Bewo融合；应用逆转录PCR（RT—PCR）检测Syncytin、Survivin基因在forskolin作用0、24h、48h、72h的绒癌细胞株BeWo中的表达：应用蛋白质印迹法检测Survivin、增殖细胞核抗原（ProliferatingCellNuclearAntigen，PCNA）蛋白在forskolin作用0、24h、48h、72h的绒癌细胞株BeWo中的表达；应用噻唑蓝比色分析实验（MTT）检测forskolin作用0、24h、48h、72h的绒癌细胞株BeWo的增殖以及绒癌细胞株BeWo融合前后对5-氟尿嘧啶（5-Fu）、甲氨蝶呤（MTX）、顺铂等不同化疗药物的敏感性。结果forskolin作用后的BeWo细胞株中Syncytin基因的表达增强，且随着forskolin作用时间的延长，Syncytin的表达越强，于48小时达到高峰；forskolin作用后的BeWo细胞株中Survivin基因的表达下降，且随着forskolin作用时间的延长，其表达SurvivinnlRNA越低；forskolin处理后的BeWo细胞株中Survivin和PCNA蛋白的表达均下降，且随着forskolin作用时间的延长，其表达越弱；③与对照组比较，forskolin处理后的BeWo细胞株的增殖能力下降，且具有统计学意义（F值分别为5．722、6．380、7．131，均P〈0．05）；随着forskolin作用时间越长，BeWo细胞株增殖能力下降越明显。同对照组比较，forskolin作用48h后的Bew0细胞株对5-Fu、MTX、顺铂等不同化疗药物的敏感性增高，且具有统计学差异（，值为5．806—11．203，均P〈0．05）。结论绒癌细胞株BeWo合体化后其增殖能力下降，且其增殖相关基因Survivin、PCNA的表达也下降。绒癌细胞株Bewo合体化后对5-Fu、MTX、顺铂等不同化疗药物的敏感性均增高，推测妊娠滋养细胞合体化可能改善恶性滋养细胞肿瘤的耐药性，但妊娠滋养细胞合体化与恶性滋养细胞肿瘤耐药的关系有待进一步研究。

Objective To develop a new therapy for gestational trophoblastic tumor （GRIT）, especially drug-resistant GTTin clinics through researching the changes in the sensitivity of BeWo before and after fusion to different chemotherapy drugs and exploring the relationship between syncytial fusion of trophoblast cells and drug resistance of GTT. Methods Forskolin was utilized to induce intercellular fusion of ehoriocarcinoma cell line BeWo. The expressions of Syncytin and Survivin mRNA in choriocarcinoma cell line BeWo treated with forskolin for 0, 24, 48 and 72 hours were detected by RT-PCR, while Western-blot was used to detect the expressions of Survivin and proliferating cell nuclear antigen （PCNA） proteins in BeWo treated with forskolin for 0, 24, 48 and 72 hours. The proliferations of choriocarcinoma cell line BeWo treated with forskolin for 0, 24, 48 and 72 hours and the sensitivity of BeWo to 5-Fu, MTX and Cisplatin before and after fusion were detected by MT＇F. Results The expression of Syncytin gene in BeWo cell line treated with forskolin increased, and with the prolonging of treatment, the expression was higher and reached its peak at 48 hour. The expression of Survivin gene in BeWo cell line treated with forskolin declined, and with the prolonging of treatment, the expression was lower. After treatment with forskolin, the expressions of Survivin and PCNA proteins in BeWo cell line declined, and as the cell line were treated longer, the expressions were lower. Compared with the control group, the proliferation of BeWo cell line treated with forskolin decreased with statistical significance （ F value was 5. 722, 6. 380 and 7. 131, respectively, all P 〈 0.05 ）. The decrease of proliferation of BeWo cell line was more serious when treatment with forskolin was longer. Compared with the control group, BeWo cell line treated with forskolin for 48 hours were more sensitive to 5-Fu, MTX and Cisplatin, and the differences were significant （ F value ranged 5. 806 - 11. 203, all P 〈 0.05 ）. Conclusion The proliferation of choriocarcinoma cell line BeWo decreases after fusion, and the expressions of related Survivin and PCNA genes in these cells also decrease. Choriocarcinoma cell line BeWo is more sensitive to 5-Fu, MTX and Cisplatin after intercellular fusion. So it can be speculated that intercellular fusion of trophoblast cells may improve drug resistance of malignant GTT. However, the relationship between intercellular fusion of trophoblast cells and drug resistance of malignant GTT needs further researches.