可用于卵巢癌诊断和术前治疗的核酸嵌合体的构建

Construction of Nucleic Acid Chimera for Diagnosis and Preoperative Treatment of Ovarian Cancer

目的:构建一种可用于卵巢癌诊断和术前治疗的核酸嵌合体。方法:设计一种癌抗原125(CA_125)-Plk1嵌合体,培养OVCAR-3卵巢癌细胞,平均分成嵌合体组(加入1μl的CA_125-Plk1嵌合体)、抗体组(加入1μl的CA_125抗体)、对照组(加入1μl的0.9%氯化钠液)。流式细胞仪检测3组结合卵巢癌细胞能力;噻唑蓝(MTT)增殖实验检测3组卵巢癌细胞增殖的情况;Tunel染色检测嵌合体组和对照组卵巢癌细胞凋亡的情况。结果:流式细胞检测结果示,嵌合体组中CA_125-Plk1嵌合体结合卵巢癌细胞的比例(83.67%)高于抗体组中CA_125抗体与卵巢癌细胞结合比例(65.12%)和对照组与卵巢癌细胞结合比例(30.74%),差异有统计学意义(P<0.05);MTT检测结果显示,嵌合体组卵巢癌细胞的增殖率(32.60%)低于对照组(62.24%)和抗体组(57.99%),差异均有统计学意义(P<0.05);Tunel染色结果显示,嵌合体组卵巢癌细胞中可以见到SiRNA荧光表达,与对照组比较,嵌合体组卵巢癌细胞凋亡比例明显上升[(78.46±6.33)%vs(19.73±4.27)%],差异有统计学意义(P<0.05)。结论:CA_125-Plk1嵌合体能特异性地结合卵巢癌细胞,通过抑制增殖和促凋亡,达到一个潜在的诊断和术前治疗作用。

Objective：To construct one nucleic acid chimera for diagnosis and preoperative treatment of ovari- an cancer. Methods：Culture of OVCAR-3 ovarian cancer cells were divided into chimerism group,antibody group and control group. A CA125-Plkl chimera was designed and PEG was used to increase the stability of nucleic acid chimera;flow cytometry method was used to detect ability of chimerism combining with ovarian cancer celIs;MTT was used to detect the influence of chimerism on proliferation of ovarian cancer cells;Tunel staining was used to detect the effect of chimerism on ovarian cancer cell apoptosis. Results： Flow cytometry showed that the chimera group（83.67%） with ovarian cancer cells was higher than that of antibody group （65. 12%）and control group （30.74%） ,the difference was statistically significant（ P 〈0.05） ;The results of MTT showed that the proliferation rate of ovarian cancer cells in the chimera group was （32.6%）, which was decreased by （62.24%）and （57.99%）, respectively （P 〈 0.05）; Tunel staining showed that compared with control group [（ 19.73±4.27） % ] ,the ovarian cancer cell apoptosis ratio in chimera group [ （78.46 ± 6.33）% ] increased significantly （P 〈0. 05）. Conclusions.We constructed a CA125-PIk1 chimera,which can specifically bind ovarian cancer cells, inhibit proliferation and promote apoptosis,reaching a potential diagnostic and preoperative therapeutic effect.