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目的 :探讨抗人乙酰肝素酶(heparanase,HPA)单克隆抗体(简称单抗)联合紫杉醇(paclitaxel,PTX)对乳腺癌MCF-7、MCF-7/F5和MDAMB-231细胞增殖、迁移和侵袭的影响。方法 :应用反转录-PCR(reverse transcription-PCR,RT-RCR)法检测抗人HPA单抗对MCF-7、MCF-7/F5和MDA-MB-231细胞HPA m RNA表达水平的影响;MTT法检测不同浓度的抗人HPA单抗、PTX或抗人HPA单抗联合PTX干预对乳腺癌细胞增殖能力的影响;FCM法、细胞划痕实验和Transwell小室法分别检测抗人HPA单抗、PTX或抗人HPA单抗联合PTX干预对乳腺癌细胞的细胞周期、细胞迁移和细胞侵袭能力的影响。结果:10和20 nmol/L抗人HPA单抗干预后,MCF-7/F5和MDAMB-231细胞中HPA m RNA的表达水平明显低于鼠Ig G组(P值均<0.05)。抗人HPA单抗和PTX单独处理后,MCF-7、MCF-7/F5和MDAMB-231细胞的增殖受到抑制,且呈剂量依赖性(P值均<0.05)。抗人HPA单抗联合PTX处理后,3组细胞的增殖抑制率均高于任一单药处理组(P值均<0.05)。抗人HPA单抗联合PTX处理后,3组细胞中G0/G1期细胞所占百分比明显低于溶剂对照组(只加培养液),G2/M期细胞所占百分比明显高于溶剂对照组(P<0.05,P<0.01),且出现亚二倍体峰。抗人HPA单抗合PTX处理后,MCF-7/F5和MDA-MB-231细胞的迁移能力和侵袭能力低于溶剂对照组、鼠Ig G组和任一单药处理组(P值均<0.05)。结论 :抗人HPA单抗联合PTX可协同抑制乳腺癌细胞的增殖、迁移和侵袭能力,其作用机制可能与二药联合作用后将细胞阻滞于G2/M期有关。
Objective: To investigate the effects of anti-human heparanase (HPA) monoclonal antibody and paclitaxel (PTX) on the proliferation and migration of breast cancer MCF-7, MCF-7 / F5 and MDAMB- Impact of invasion. Methods: The effect of anti-human HPA McAb on the expression of HPA m RNA in MCF-7, MCF-7 / F5 and MDA-MB-231 cells was detected by reverse transcription-PCR (RT-RCR) The effects of different concentrations of anti-human HPA monoclonal antibody, PTX or anti-human HPA monoclonal antibody combined with PTX on the proliferation of breast cancer cells were detected by MTT assay. Anti-human HPA monoclonal antibody was detected by FCM assay, cell scratch assay and Transwell chamber assay, Effects of PTX or anti-human HPA monoclonal antibody combined with PTX on the cell cycle, cell migration and cell invasiveness of breast cancer cells. Results: The expression of HPA m RNA in MCF-7 / F5 and MDAMB-231 cells was significantly lower than that in the murine Ig G group (P <0.05) after intervention with 10 and 20 nmol / L anti-human HPA monoclonal antibody. The proliferation of MCF-7, MCF-7 / F5 and MDAMB-231 cells was inhibited in a dose-dependent manner (all P <0.05) after treatment with anti-human HPA mAb and PTX alone. Anti-human HPA McAb combined with PTX treatment, the proliferation inhibition rate of three groups were higher than any single drug treatment group (P <0.05). The percentage of G0 / G1 phase cells in 3 groups of cells was significantly lower than that of solvent control group (only medium), the percentage of cells in G2 / M phase was significantly higher than that in solvent control group ( P <0.05, P <0.01), and sub-diploid peak. The migration ability and invasion ability of MCF-7 / F5 and MDA-MB-231 cells after anti-human HPA monoclonal antibody treatment with PTX were lower than those of solvent control group, mouse Ig G group and any single drug treatment group (P < 0.05). CONCLUSION: Anti-human HPA monoclonal antibody combined with PTX can synergistically inhibit the proliferation, migration and invasion of breast cancer cells. The mechanism may be related to the cell arrest in G2 / M phase after combination of two drugs.