-
摘要:
目的 探讨白喉毒素（diphtheria toxin，DT）片段DT390与靶向肽TMTP1的融合蛋白对卵巢癌的治疗效果。 方法 以卵巢癌顺铂耐药细胞株C13*及顺铂敏感细胞株OV2008为细胞模型，设对照组、TMTP1组、DT390-TMTP1组、DT390-biTMTP1组及DT390-triTMTP1组。激光共聚焦显微镜观察各组细胞核的形态。MTT法检测细胞存活率，流式细胞术检测细胞凋亡率。建立裸鼠C13*细胞皮下瘤模型，观察肿瘤的形成及生长，TUNEL法检测皮下瘤组织的细胞凋亡率。 结果 激光共聚焦显微镜观察到DT390-biTMTP1和DT390-triTMTP1引起细胞核皱缩和碎裂。MTT结果显示细胞存活率随着DT390-biTMTP1与DT390-triTM-TP1浓度增加而显著降低。流式细胞术检测结果显示DT390-biTMTP1组与DT390-triTMTP1组的细胞凋亡率显著增加（P＜0.05），并且DT390-biTMTP1组与DT390-triTMTP1组的C13*细胞凋亡率为66.0%±12.0%与72.9%±4.6%，较OV2008细胞的55.5%±8.9%与65.1%±9.8%更高。裸鼠皮下瘤结果显示DT390-biTMTP1与DT390-triTMTP1均显著抑制卵巢癌皮下瘤的形成（P＜0.01）及生长（P＜0.05）。TUNEL检测显示DT390-biTMTP1组和DT390-triTMTP1组皮下瘤组织的细胞凋亡率显著增强（P＞0.05）。同时，以上实验结果均显示DT390-TMTP1对C13*及OV2008细胞无明显作用（P＞0.05）。 结论 DT390-biTMTP1及DT390-triTMTP1融合蛋白靶向抑制卵巢癌细胞，显示了潜在的临床应用前景。 Abstract:Objective To investigate targeted therapy of ovarian cancer with new fusion proteins that were produced by fusing the first 390 amino acids of diphtheria toxin (DT390) to the TMTP1 peptide. Methods The cisplatin-resistant cell line, C13*, and cisplatin-sensitive cell line, OV2008, were selected as models and divided into control, TMTP1, DT390-TMTP1, DT390-biTMTP1, and DT390-triTMTP1 groups. Laser scanning confocal microscopy was used to observe nuclear morphology. 3-(4, 5-Dimethylthiazol-2-yl) -2, 5-diphenyltetrazolium bromide (MTT) and flow cytometry assays were used to detect cell survival and apoptosis, respectively. The formation of subcutaneous tumors in nude mice following injection of C13* cells was used to observe the formation and growth of ovarian cancer. Apoptosis of cells in the subcutaneous tumor tissue was detected by the terminal deoxynucleotidyl transferase dUTP nick-end labeling (TUNEL) assay. Results Laser scanning confocal microscopy showed that DT390-biTMTP1 and DT390-triTMTP1 induced nuclear shrinkage and fragmentation. The MTT assay showed that cell survival was obviously reduced with increasing concentrations of DT390-biTMTP1 and DT390-triTMTP1. Flow cytometry revealed that DT390-biTMTP1 and DT390-triTMTP1 significantly increased cell apoptosis (P < 0.05). The apoptosis rates of the DT390-biTMTP1 and DT390-triTMTP1 groups were 66.0%±12.0% and 72.9%±4.6%, respectively. These were higher than the 55.5%±8.9% and 65.1%±9.8% obvserved in OV2008 cells. DT390-biTMTP1 and DT390-triTMTP1 significantly inhibited the tumor formation (P < 0.01) and growth (P < 0.05), and increased apoptosis (P < 0.05) of subcutaneous tumors. However, DT390-TMTP1 had insignificant effects on C13* and OV2008 cells. Conclusions DT390-biTMTP1 and DT390-triTMTP1 preferentially targeted and inhibited ovarian cancer cells. These fusion proteins may be a promising strategy for clinical therapy of ovarian cancer. -
Key words:
- DT390 /
- TMTP1 /
- ovarian cancer /
- targeted therapy
-
[1] 严丽梅, 李秀琴.2017肿瘤生物治疗最新进展暨"卵巢癌规范化、个体化、精准治疗高峰论坛"纪要[J].中国实用妇科与产科杂志, 2017, 33(8):874-880. http://kns.cnki.net/KCMS/detail/detail.aspx?filename=zgsf201708034&dbname=CJFD&dbcode=CJFQ [2] Legge F, Ferrandina G, Salutari V, et al. Biological characterization of ovarian cancer: prognostic and therapeutic implications[J]. Ann Oncol, 2005, 16 (Suppl 4):iv95-101. http://citeseerx.ist.psu.edu/viewdoc/summary?doi=10.1.1.555.5205 [3] 申薇, 梁冰锋, 李秀荣, 等.卵巢癌耐药细胞株SKOV3/DDP的建立及其与凋亡途径蛋白的关系[J].河北医科大学学报, 2014, 35(10): 1135-1139. doi: 10.3969/j.issn.1007-3205.2014.10.007 [4] Tse BW, Collins A, Oehler MK, et al. Antibody-based immunotherapy for ovarian cancer: where are we at[J]? Ann Oncol, 2014, 25(2): 322-331. doi: 10.1093/annonc/mdt405 [5] Torchilin VP. Drug targeting[J]. Eur J Pharm Sci, 2000, 11 (Suppl 2): S81-91. http://europepmc.org/abstract/MED/11033430 [6] Ma X, Lv P, Ye S, et al. DT390-triTMTP1, a novel fusion protein of diphtheria toxin with tandem repeat TMTP1 peptide, preferentially targets metastatic tumors[J]. Mo Pharm, 2013, 10(1):115-126. doi: 10.1021/mp300125k [7] 叶双梅. DT390与TMTP1融合毒素的构建及其靶向治疗高转移性肿瘤的研究[D]. 武汉: 华中科技大学, 2011. [8] Benito V, Lubrano A, Arencibia O, et al. Laparoscopic extraperitoneal para-aortic lymphadenectomy in the staging of locally advanced cervical cancer: is it a feasible procedure at a peripheral center[J]? Int J Gynecol Cancer, 2012, 22(2):332-336. doi: 10.1097/IGC.0b013e31823c241b [9] Gatalica Z, Lele SM, Rampy BA, et al. The expression of Fhit protein is related inversely to disease progression in patients with breast carcinoma[J]. Cancer, 2000, 88(6):1378-1383. doi: 10.1002/(ISSN)1097-0142 [10] Huo YR, Richards A, Liauw W, et al. Hyperthermic intraperitoneal chemotherapy (HIPEC) and cytoreductive surgery (CRS) in ovarian cancer: A systematic review and meta-analysis[J]. Eur J Surg Oncol, 2015, 41(12):1578-1589. doi: 10.1016/j.ejso.2015.08.172 [11] 胡秋月, 余祖江.分子靶向治疗药物在原发性肝癌中的研究进展[J].国际消化病杂志, 2017, 37(1):20-23. http://med.wanfangdata.com.cn/Paper/Detail/PeriodicalPaper_sxzlyx201408078 [12] 张景锋, 朱宽佑, 徐秋良, 等.白喉毒素作用机制及其应用的研究进展[J].畜牧与饲料科学, 2017, 38(10):33-37. http://d.old.wanfangdata.com.cn/Periodical/nmgxmkx201710010 [13] Lansigan F, Stearns DM, Foss F. Role of denileukin diftitox in the treatment of persistent or recurrent cutaneous T-cell lymphoma[J]. Cancer Manag Res, 2010, 2:53-59. http://europepmc.org/articles/PMC3004568/ [14] Landon LA, Zou J, Deutscher SL. Is phage display technology on target for developing peptide-based cancer drugs[J]? Curr Drug Discov Technol, 2004, 1(2):113-132. http://europepmc.org/abstract/med/16472251 [15] Yang W, Luo D, Wang S, et al. TMTP1, a novel tumor-homing peptide specifically targeting metastasis[J]. Clin Cancer Res, 2008, 14(17):5494-5502. doi: 10.1158/1078-0432.CCR-08-0233 -
下载:
点击查看大图
图(4)
计量
- 文章访问数: 87
- HTML全文浏览量: 49
- PDF下载量: 6
- 被引次数: 0
