基础医学与临床 ›› 2018, Vol. 38 ›› Issue (10): 1438-1442.

• 研究论文 • 上一篇    下一篇

硼替佐米促进霍奇金淋巴瘤细胞系L428的凋亡

陈宛丽,关红梅,王舒   

  1. 南阳市第二人民医院
  • 收稿日期:2017-10-27 修回日期:2018-02-01 出版日期:2018-10-05 发布日期:2018-09-28
  • 通讯作者: 陈宛丽 E-mail:2133056357@qq.com

Bortezomib promotes cell apoptosis in Hodgkin lymphoma cell line L428

  • Received:2017-10-27 Revised:2018-02-01 Online:2018-10-05 Published:2018-09-28

摘要: 目的 探究硼替佐米不同剂量对L428细胞株形态学、生长抑制率、细胞凋亡率以及肿瘤细胞侵袭转移的影响,并阐明硼替佐米影响淋巴瘤侵袭转移的机制,为临床上经典型霍奇金淋巴瘤患者的治疗提供新的思路和靶点。方法 采用研究级倒置显微镜观察加入10 nM硼替佐米培养24 h后,L428细胞的生长状态;采用MTT法检测检测不同浓度硼替佐米处理不同时程后,L428细胞的生长抑制率;采用TUNEL试剂盒检测不同浓度硼替佐米处理不同时程后,L428细胞的凋亡率;采用Transwell法检测不同浓度的硼替佐米对于肿瘤细胞L428侵袭、转移能力的影响;采用western blot法检测硼替佐米对凋亡相关蛋白Bax和Bcl-2的影响。结果 与空白对照组相比,加入10 nM硼替佐米培养24 h后,细胞状态不佳,出现拉丝、稀疏等现象;MTT结果显示,随着硼替佐米浓度由10 nM增至50 nM,处理时间由12 h增至36 h,细胞生长抑制率呈升高趋势(P<0.05);TUNEL结果显示,随着硼替佐米浓度由10 nM增至50 nM,处理时间由24h增至72h,细胞凋亡率呈升高趋势(P<0.05);Transwell结果表明,随着硼替佐米浓度由10 nM增至50 nM,肿瘤细胞的侵袭转移能力逐渐减弱(P<0.05);western blot结果显示,在L428细胞株中加入10 nM、30 nM、50 nM硼替佐米培养48 h后,Bcl-2蛋白水平呈浓度依赖性下调趋势(P<0.05);Bax蛋白水平呈浓度依赖性上调趋势(P<0.05)。结论 硼替佐米可以通过影响凋亡蛋白的表达来影响肿瘤细胞的生长、侵袭、转移能力,且在一定用药剂量范围里对肿瘤细胞的抑制作用呈浓度依赖性,对临床上淋巴瘤患者的治疗具有重要的参考价值。

关键词: 硼替佐米, 经典型霍奇金淋巴瘤, 生长抑制率, 凋亡率, 侵袭转移能力

Abstract: Objective: To explore the influence on L428 cell lines morphology, growth inhibition rate, apoptosis rate, tumor invasion and metastasis of different doses of Bortezomib, and then illustrate the mechanism that how Bortezomib influence the tumor invasion and metastasis so that we can provide new thought and therapeutic target for the clinical treatment of lymphoma. Methods: Inverted microscope was used to observe L428 cell growth state after treating cells with 10 nM Bortezomib for 24 h; MTT was used to detect the L428 cell growth inhibition rate after treating cells with different Bortezomib concentration and time; TUNEL kit was used to detect the L428 cell apoptosis rate after treating cells with different Bortezomib concentration and time; Transwell was carried out to detect the invasion and metastasis of lymphoma influenced by different Bortezomib doses; Western blot was carried to detect the apoptosis proteins expression of Bcl-2 and Bax influenced by different Bortezomib doses. Results: Cells have appeard cliques, wire drawing and sparse phenomenon after treating with 10nM Bortezomib for 24h when compared with the blank control group; MTT data has indicated that as the Bortezomib concentration increased from 10nM to 50nM and the treating time increased from 12h to 36h, ?the cell growth inhibition rate showed a trend of rising (P<0.05); TUNEL data has indicated that as the Bortezomib concentration increased from 10nM to 50nM and the treating time increased from 24h to 72h, ?the cell apoptosis rate showed a trend of rising (P<0.05); Transwell has indicated that as the Bortezomib concentration increased from 10nM to 50nM, the?cell invasion and metastasis ability decreased gradually (P<0.05); ?The Bcl - 2 protein level showed a trend of concentration dependence downgrade (P<0.05) and Bax protein level showed a trend of concentration dependence upgrade (P<0.05) when treating cells with 10nM, 30nM, 50nM Bortezomib for 48h. Conclusions: Bortezomib can influence the growth state and the ability of invasion and metastasis of tumor cells by influencing the expression of apoptosis proteins, and the inhibition rate of tumor cells are concentration dependence?in a certain dosage range, the study has important reference value for clinical treatment of lymphoma.

Key words: Bortezomib, lymphoma, growth inhibition rate, apoptosis rate, invasion and metastasis ability