表达特异性多肽的噬菌体对卵巢癌细胞生物学行为的影响

Effects of selected phages with specific peptide ligands on the biological behaviors of ovarian cancer cells

目的探讨表达特异性多肽的噬菌体对卵巢癌细胞生物学行为的影响。方法选取卵巢癌细胞株A2780、SKOV3,分别与特异性的噬菌体共同作用,通过台盼蓝拒染法、流式细胞仪、平板克隆形成实验、四甲基偶氮唑蓝比色法、Boyden小室体外侵袭实验等方法,检测卵巢癌细胞生物学行为的变化。将A2780、SKOV3细胞按实验目的分别分为3组:(1)实验组,加入可与卵巢癌细胞特异性结合的噬菌体(阳性噬菌体);(2)空白对照组,不加噬菌体;(3)阴性对照组,加入不与卵巢癌细胞结合的噬菌体(阴性噬菌体)。结果实验组、阴性对照组和空白对照组的平均活细胞比例分别为(72·1±6·2)%、(84·8±4·6)%和(93·1±2·5)%,与阴性对照组及空白对照组比较,差异均有统计学意义(P<0·05);实验组A2780和SKOV3细胞的凋亡率分别为20·39%和43·99%,而阴性对照组均未见凋亡峰的出现;实验组细胞的平均克隆形成率为4%,阴性对照组为15%,空白对照组为15%,实验组与阴性对照组及空白对照组分别比较,差异均有统计学意义(P<0.05);实验组A2780和SKOV3细胞的生长抑制率分别为11·07%和9·58%,均明显高于各自的阴性对照组(分别为2·05%和1·09%),与各自的阴性对照组比较,差异有统计学意义(P<0.05);实验组A2780和SKOV3细胞的侵袭指数,都低于各自的阴性对照组和空白对照组,分别与各自的阴性对照组和空白对照组比较,差异均有统计学意义(P<0·05)。结论筛选出的特异性噬菌体对卵巢癌细胞的生长、增殖和迁移有一定的下调作用,与噬菌体结合的卵巢癌细胞表面受体可能与肿瘤的发生和转移有关。

Objective To evaluate the effects of the selected phages with the specific peptide ligands upon the biological behaviors of the ovarian cancer cells. Methods Two ovarian cancer cell lines, A2780 and SKOV3, were used in the study. They were divided into three groups respectively： study group, blank control group and negative control group. The effects of the phages were evaluated by trypan blue staining, flow cytometry, colony formation test, methyl thiazolyl tetrazolium （MTT） assay and Boyden chamber invasion assay. Results The average viability of ovarian cancer cells of study groups was （72. 1 ± 6. 2） %, higher than that of negative control group （ 84. 8 ± 4. 6, P 〈 0. 05 ） ; the apoptosis ratio of the A2780 and SKOV3 cells of study groups increased （20. 39% and 43.99% vs 0） ; the average colony formation rate of the ovarian cancer cells was of study groups 4% , lower than the control group （ 15% , P 〈 0.05 ） ; the inhibition rates of cell proliferation of A2780 and SKOV3 cells of study groups were 11.07% and 9. 58% , significantly higher than that of their respective negative control （ 2.05% and 1.09% ） ; the invasion index of the ovarian cancer cells decreased compared with the control. Conclusion The selected phages downregulate the growth, proliferation and invasion ability of the ovarian cancer cells to certain extent, the identified peptide ligands may play a role in tumorigenesis and metastatic transformation of ovarian cancers.