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目的:探讨靶向小鼠肝细胞再生磷酸酶-3(mouse phosphatase of regenerating liver-3,mPRL-3)的DNA疫苗对小鼠乳腺癌D2F2细胞体内生长的抑制作用。方法:构建靶向mPRL-3的真核表达载体pVAX1-mPRL-3,并转染至鹌鹑肌成纤维细胞QM7内,Western blotting检测mPRL-3蛋白在QM7细胞中的表达;通过重组慢病毒Lv-mPRL-3或对照载体Lv-Ctrl感染小鼠乳癌D2F2细胞,分别建立高表达mPRL-3的mPRL-3-D2F2细胞或对照NC-D2F2细胞,Western blotting检测小鼠乳腺癌D2F2细胞中mPRL-3蛋白的表达。BALB/c小鼠左侧乳腺脂肪垫下分别接种mPRL-3-D2F2和NC-D2F2细胞后,通过基因枪接种pVAX1-mPRL-3疫苗(mPRL-3-D2F2/pVAX1-mPRL-3),同时设立疫苗对照组(mPRL-3-D2F2/pVAX1-Ctrl)和细胞对照组(NC-D2F2/pVAX1-mPRL-3),检测小鼠的肿瘤体积及生存期。结果:pVAX1-mPRL-3质粒经酶切鉴定及测序验证均正确,并能在QM7细胞中表达。Western blotting检测结果显示,慢病毒感染的mPRL-3-D2F2细胞中mPRL-3蛋白过表达,而NC-D2F2细胞中不表达mPRL-3蛋白。荷瘤小鼠经pVAX1-mPRL-3 DNA疫苗免疫,其肿瘤体积明显低于对照组[(835.3±509.8)vs(1 543.0±578.4)mm3,P<0.01],且pVAX1-mPRL-3疫苗能显著延长荷瘤小鼠的生存期(中位生存期55.5 vs 38 d,P<0.05)。结论:基因枪接种的靶向mPRL-3的DNA疫苗能抑制高表达mPRL-3的小鼠乳腺癌D2F2细胞移植瘤的生长,并延长荷瘤小鼠生存期,提示其对mPRL-3阳性肿瘤有潜在的治疗作用。
AIM: To investigate the inhibitory effect of mouse DNA vaccine targeting mouse phosphatase of regenerating liver-3 (mPRL-3) on the growth of mouse breast cancer D2F2 cells in vivo. Methods: The eukaryotic expression vector pVAX1-mPRL-3 targeting mPRL-3 was constructed and transfected into quail myofibroblasts QM7. The expression of mPRL-3 protein in QM7 cells was detected by Western blotting. The recombinant lentivirus Lv The mPRL-3-D2F2 cells or the control NC-D2F2 cells with high mPRL-3 expression were established by inoculation of mPRL-3 or control vector Lv-Ctrl into D2F2 cells. Western blotting was used to detect mPRL- 3 protein expression. BALB / c mice were inoculated with mPRL-3-D2F2 and NC-D2F2 cells respectively on the left mammary fat pad, and the pVAX1-mPRL-3 vaccine (mPRL-3-D2F2 / pVAX1-mPRL-3) The vaccine control group (mPRL-3-D2F2 / pVAX1-Ctrl) and the cell control group (NC-D2F2 / pVAX1-mPRL-3) were established to test the tumor volume and survival of mice. Results: Plasmid pVAX1-mPRL-3 was verified by restriction enzyme digestion and sequencing, and was able to express in QM7 cells. Western blotting showed that mPRL-3 protein was overexpressed in lentivirus-infected mPRL-3-D2F2 cells but not mPRL-3 protein in NC-D2F2 cells. The tumor volume of tumor-bearing mice immunized with pVAX1-mPRL-3 DNA vaccine was significantly lower than that of the control group [(835.3 ± 509.8) vs (5443.0 ± 578.4) mm3, P <0.01] Significantly prolong the survival of tumor-bearing mice (median survival 55.5 vs 38 days, P <0.05). CONCLUSION: The DNA vaccine targeting mPRL-3 administered by gene gun can inhibit the growth of xenografted breast cancer D2F2 cells that have mPRL-3 overexpression and prolong the survival of tumor-bearing mice, suggesting that mPRL-3 positive tumors Potential therapeutic effect.