摘要
目的 探讨β-榄香烯对分化型甲状腺癌细胞TPC-1131碘(I)敏感性的影响及相关机制。方法 将TPC-1细胞随机分为对照组、131I组、β-榄香烯组和131I+β-榄香烯组。采用细胞计数试剂盒-8(CCK-8)实验检测TPC-1细胞存活率,克隆形成实验检测TPC-1细胞克隆形成数,流式细胞术检测TPC-1细胞凋亡率,Hoechst 33258染色检测TPC-1细胞凋亡形态,Transwell检测TPC-1细胞迁移水平,蛋白免疫印迹(Western blot)检测TPC-1细胞细胞周期蛋白D1(CyclinD1)、细胞周期蛋白依赖性激酶抑制剂1A(p21)、B淋巴细胞瘤-2相关X蛋白(Bax)、B淋巴细胞瘤-2(Bcl-2)、基质金属蛋白酶-2(MMP-2)、基质金属蛋白酶-9(MMP-9)、p-p65、p65、磷酸化核因子κB抑制蛋白α(p-IκBα)和核因子κB抑制蛋白α(IκBα)蛋白表达水平。结果 对照组、131I组、β-榄香烯组、131I+β-榄香烯组的细胞存活率依次是100.00%±0.00%、83.69%±15.13%、91.35%±14.30%和56.13%±9.05%,克隆形成数依次是507.19±72.31、421.25±65.43、453.30±79.80和317.71±53.24。与对照组比较,131I组TPC-1细胞存活率、克隆形成数、迁移数和CyclinD1、Bcl-2、MMP-2、MMP-9水平明显降低(P<0.05),凋亡率和p21、Bax、p-p65/p65、p-IκBα/IκBα水平明显增加(P<0.05),与131I组比较,131I+β-榄香烯组TPC-1细胞存活率、克隆形成数、迁移数和CyclinD1、Bcl-2、MMP-2、MMP-9、p-p65/p65、p-IκBα/IκBα水平明显降低(P<0.05),凋亡率和p21、Bax水平明显增加(P<0.05)。对照组和β-榄香烯组细胞核呈均匀的淡蓝色,131I组和131I+β-榄香烯组细胞核出现碎裂、浓缩、染色呈亮蓝色等凋亡特征,且131I+β-榄香烯组的凋亡特征更明显。结论 榄香烯可提高TPC-1细胞131I敏感性,其机制可能与β-榄香烯抑制131I引起的核因子κB(NF-κB)信号通路激活有关。
Objective To investigate the effect of β-elemene on131I sensitivity of differentiated thyroid cancer cell TPC-1 and its related mechanism. Methods TPC-1 cells were randomly divided into control group,131I group, β-elemene group and131I+β-elemene group. Cell counting Kit 8(CCK-8) assay was used to detect the survival rate of TPC-1 cells. Clonal formation assay was used to detect the clonal formation number of TPC-1 cells. Flow cytometry was used to detect the apoptosis rate of TPC-1 cells. Hoechst 33258 staining was used to detect the apoptotic morphology of TPC-1 cells. Transwell was used to detect the cell migration level of TPC-1 cells. Western blot was used to detect the protein expression level of CyclinD1, cyclin-dependent kinase inhibitors 1A(P21), B-lymphocytoma-2-associated X protein(Bax), B-lymphocytoma-2(Bcl-2), matrix metalloproteinase-2(MMP-2), matrix metalloproteinase-9(MMP-9), p-p65, p65, phosphorylated nuclear factor κB inhibitory protein α(p-IκBα), nuclear factor κB inhibitory protein α(IκBα) of TPC-1 cells. Results The cell survival rates of control group,131I group, β-elemene group and131I+β-elemene group were 100.00%±0.00%, 83.69%±15.13%, 91.35%±14.30% and 56.13%±9.05%, respectively. The number of clone formation was 507.19±72.31, 421.25±65.43, 453.30±79.80 and 317.71±53.24, respectively. Compared with the control group, the survival rate, number of clone formation, number of migration, levels of CyclinD1, Bcl-2, MMP-2 and MMP-9 of TPC-1 cells in the131I group were significantly decreased(P<0.05), while the apoptosis rate, p21, Bax, p-p65/p65 and p-IκBα/IκBα levels were significantly increased(P<0.05). Compared with the131I group, the survival rate, number of clone formation, number of migration, levels of CyclinD1, Bcl-2, MMP-2, MMP-9, p-p65/p65, p-IκBα/IκBα of TPC-1 cells in the131I+β-elemene group were significantly decreased(P<0.05), while the apoptosis rate, p21, Bax levels were significantly increased(P<0.05). The cell nucleus of control group and β-elemene
作者
张雅兰
康柳枝
黄培瑜
陈文发
ZHANG Ya-lan;KANG Liu-zhi;HUANG Pei-yu;CHEN Wen-fa(The Second Affiliated Hospital of Fujian Medical University,Quanzhou Fujian 362000,China)
出处
《毒理学杂志》
CAS
2023年第1期24-30,共7页
Journal of Toxicology
基金
福建医科大学启航基金(2019QH1127)。
