鸦胆子油乳及联合顺铂对卵巢癌SKOV3裸鼠移植瘤生长抑制的影响

Effect of Brucea javanica Fruit Oil Emulsion Combined Cisplatin on the Growth Inhibition of Transplanted Tumor in Human Ovarian Cancer SKOV3 Nude Mice: an Experimental Study

目的研究鸦胆子油乳对人卵巢癌细胞系SKOV3细胞及SKOV3裸鼠移植瘤的生长抑制作用。方法 MTT法测定不同浓度鸦胆子油乳单独及联合顺铂对人卵巢癌细胞SKOV3的生长抑制作用。采用人卵巢癌细胞SKOV3移植方法构建卵巢癌裸鼠模型,将32只卵巢癌裸鼠随机分为4组,每组8只。空白对照组(简称对照组)、鸦胆子油乳组(简称鸦胆子组)、鸦油顺铂联合组(简称联合组)及顺铂组。A、B组分别每隔2日腹腔注射生理盐水、鸦胆子油乳注射液0.2 m L/20 g,联合组第1天腹腔注射顺铂0.2 m L(3 mg/kg),第2天腹腔注射0.2 m L/20 g鸦胆子油乳注射液,D组每隔2日腹腔注射顺铂0.2 m L(3 mg/kg)。各组均注射6次,末次腹腔注射后48 h断颈处死裸鼠,观察腹腔肿瘤组织病灶形成情况,取肿瘤病灶标本进行HE染色和Western blot检测肿瘤组织运动相关蛋白MRP-1/CD9和整合素α-5表达水平。结果鸦胆子油乳对人卵巢癌SKOV3细胞的抑制作用呈剂量—时间依赖关系,鸦胆子油乳与顺铂联合应用对人卵巢癌SKOV3细胞的抑制作用显著高于这两种药物的单独应用。Western blot结果显示鸦胆子组和顺铂组MRP-1/CD9和整合素α-5的表达均上调,且差异有统计学意义(P<0.05),但联合组表达下调,差异有统计学意义(P<0.05)。结论鸦胆子油乳注射液腹腔给药治疗卵巢癌具有可行性和一定的疗效性,并且鸦胆子油乳可能以MRP-1/CD9和整合素α-5为靶点抑制肿瘤细胞侵袭和转移,但具体的抗肿瘤机制尚未明确。

Objective To study the growth inhibition effect of Brucea javanica Fruit Oil Emulsion（ BJFOE） on human ovarian caner SKOV3 cells and the transplanted tumor of SKOV3 nude mice. Methods Growth inhibition effects of different concentrations BJFOE alone or its combination with cisplatin on human ovarian cancer cell SKOV3 were measured using MTT method. The orthotopic transplantation tumor model of human ovarian cancer SKOV3 cell lines was established in nude mice. Totally 32 ovarian cancer nude mice were randomly divided into 4 groups,i. e.,the blank control group（ Group A）,the BJFOE group（ Group B）,the BJFOE combined Cisplatin group（ Group C）,and the Cisplatin control group（ Group D）,8 in each group. Mice in Group A were intraperitoneally injected with normal saline（ 0. 2 m L /20 g）,once per two days. Mice in Group B were intraperitoneally injected with BJFOE（ 0. 2 m L /20 g）,once per two days. Mice in Group C were intraperitoneally injected with cisplatin（ 3 mg / kg） 0. 2 m L on the first day,and intraperitoneally injected with BJFOEon the second day. Mice in Group D were intraperitoneally injected with cisplatin（ 3 mg / kg） 0. 2 m L,once per two days.All mice were injected for six times,and sacrificed 48 h after the last injection. The lesion formation of the abdominal tumor tissue was observed. Tumor specimens were obtained to perform HE staining. Expression levels of MRP-1 / CD9 and integrinα-5 were detected using Western blot. Results The inhibition of BJFOE was time-dose dependently correlated with its inhibition effect of SKOV3 cells. The inhibition effect of BJFOE in combination of cisplatin was significantly superior to that of using any of the two drugs alone. Western blot results showed expression levels of MRP-1 / CD9 and integrinα-5 were up-regulated in Group B and Group D with statistical difference（P 〈 0. 05）. But they were down-regulated in Group C with statistical difference（P 〈 0. 05）. Conclusions Intraperitoneal injecting BJFOE was feasible and effective for treating ovarian cancer. BJFOE also could inhibit the invasion and migration of tumor cells targeting at MRP-1 / CD9 and integrinα-5. But its specific anti-tumor mechanism was not clearly probed.