下调AFAP-1L2表达增强顺铂对子宫内膜癌细胞的杀伤作用

Down-regulating AFAP-1L2 attenuates the killing effect of cisplatin to endometrial cancer cells

目的观察AFAP-1L2下调后对顺铂干预的子宫内膜癌KLE细胞增殖、凋亡及细胞周期的影响,为子宫内膜癌治疗提供新的靶点。方法以Western blot及q RT-PCR检测不同分化程度子宫内膜癌细胞(KLE,Ishikawa)中AFAP-1L2蛋白及m RNA表达;构建siAFAP-1L2质粒并转染子宫内膜癌细胞,检测转染效率;顺铂干预siNegative Control细胞为对照组(DDP组),对siAFAP-1L2干扰细胞(siAFAP-1L2+DDP组)给予不同浓度顺铂干预或同一浓度不同培养时间下以MTT法观察细胞存活率,流式细胞术观察细胞凋亡率,PI法观察细胞周期。结果 AFAP-1L2蛋白及m RNA在低分化KLE细胞株中表达水平明显高于高分化Ishikawa细胞株。AFAP-1L2下调后能够增加KLE细胞对DDP化疗敏感度,并具有剂量依赖性及时间依赖性。下调AFAP-1L2表达能够促进DDP干预的KLE细胞凋亡,使更多细胞生长停滞。结论 siAFAP-1L2沉默可增强顺铂对子宫内膜癌细胞的杀灭作用,为子宫内膜癌治疗研究的候选靶点。

Objective To observe the effect of AFAP-1 L2 down-regulating on the proliferation, apoptosis and cell cycle of endometrial cancer cells （ KLE, Ishikawa ） interfered by eisplatin. Methods Western blot and （qRT）-PCR were used to detect AFAP-1L2 protein and mRNA expressions in different differentiation endometrial cancer cells. The siAFAP-1L2 plasmid was constructed and transfected into endometrial cancer cells. The cell viability was observed by MTT assay in different DDP concentration or different culture time in siAFAP-1L2 interfering eells（siAFAP-1L2＋DDP group）. Cell apoptosis and cycle were detected by flow cytometry and PI method. Results The expressed level of AFAP-1L2 protein and mRNA was higher in low differentiation KLE cells than in high differentiation Ishikawa cells. AFAP-1L2 silence attenuated the sensitivity of KLE cells to DDP chemotherapy with concentration dependent and time dependent, attenuated KLE cell apoptosis deah with DDP and more cells growing stop. Conclusion siAFAP-1L2 interfering may attenuate the killing effect of cisplatiu to endometrial cancer cells and may become a candidate target for endometrial cancer therapeutic research.