Objective: To explore the effect of carvacrol on the biological behavior of leukemia cells and its regulation to circ-0008717/miR-217 molecular axis.
Methods: Human acute lymphoblastic leukemia cells Molt-4 were cultured in vitro, and different concentrations of carvacrol were added to the cells. si-NC and si-circ-0008717 were transfected into Molt-4 cells (si-NC group, si-circ-0008717 group). pcDNA, pcDNA-circ-0008717, anti-miR-NC, anti-miR-217 were transfected into Molt-4 cells and then added to carvacrol-treated cells (carvacrol+pcDNA group, carvacrol+pcDNA-circ-0008717 group, carvacrol+anti-miR-NC group, carvacrol+anti-miR-217 group). MTT, plate clone formation experiment, and flow cytometry were used to detect the viability of the cell, colony formation number, and apoptosis rate of cells, respectively. The RT-qPCR method was used to detect the expression levels of circ-0008717 and miR-217. The dual luciferase reporter gene experiment was used to detect the targeting relationship between circ-0008717 and miR-217.
Results: After carvacrol treatment, the cell viability decreased significantly (r=-0.9405), expression level of circ-0008717 decreased (r=-0.9117), colonies formed number decreased (r=-0.9256), while the cell apoptosis rate increased (r= 0.8464), and the expression level of miR-217 increased (r=0.9468). Compared with the si-NC group, the expression level of miR-217 in si-circ-0008717 group increased (P＜0.001), the cell apoptosis rate increased (P＜0.001), while cell viability decreased (P＜0001), the number of colonies formed decreased (P＜0.001). Compared with the carvacrol+pcDNA group, the cell viability of the carvacrol+pcDNA-circ-0008717 group increased (P＜0.001), the number of colonies formed increased (P＜0.001), while the cell apoptosis rate decreased (P＜0.001). circ-0008717 could target miR-217. The cell viability of the carvacrol+anti-miR-217 group increased (P＜0.001), and the number of colonies formed increased (P＜0.001), while the cell apoptosis rate decreased (P＜0001) as compared with the carvacrol+anti-miR-NC group.
Conclusion: Carvacrol can promote the expression of miR-217 by down-regulating the expression of circ-0008717, thereby reducing the proliferation and cloning ability of leukemia cells and promoting cell apoptosis.
题目: 香芹酚对白血病细胞生物学行为的影响及其机制.
目的: 探讨香芹酚对白血病细胞生物学行为的影响及其对circ-0008717/miR-217分子轴的调控作用.
方法: 体外培养人急性淋巴细胞白血病细胞Molt-4，分别加入不同浓度的香芹酚处理细胞；si-NC、si-circ-0008717分别转染入Molt-4细胞（si-NC组、si-circ-0008717组），pcDNA、pcDNA-circ-0008717、anti-miR-NC、anti-miR-217分别转染入Molt-4细胞后加入香芹酚处理细胞（香芹酚+pcDNA组、香芹酚+pcDNA-circ-0008717组、香芹酚+anti-miR-NC组、香芹酚+anti-miR-217组）；MTT、平板克隆形成实验、流式细胞术分别检测细胞活力、集落形成数、细胞凋亡率；RT-qPCR法检测circ-0008717和miR-217的表达量；双荧光素酶报告基因实验检测circ-0008717与miR-217的靶向关系.
结果: 经香芹酚处理后，细胞活力明显降低（r=-0.9405），circ-0008717的表达水平明显降低（r=-0.9117），集落形成数明显减少（r=-0.9256），细胞凋亡率明显升高（r=0.8464），miR-217的表达水平明显升高（r=0.9468）；与si-NC组比较，si-circ-0008717组miR-217的表达水平升高（P＜0.001），细胞活力降低（P＜0001），集落形成数明显减少（P＜0.001），细胞凋亡率升高（P＜0.001）；与香芹酚+pcDNA组比较，香芹酚+pcDNA-circ-0008717组细胞活力升高（P＜0.001），集落形成数明显增多（P＜0.001），细胞凋亡率明显降低（P＜0001）；circ-0008717可靶向调控miR-217的表达；与香芹酚+anti-miR-NC组比较，香芹酚+anti-miR-217组细胞活力明显升高（P＜0.001），集落形成数明显增多（P＜0.001），细胞凋亡率明显降低（P＜0.001）.
结论: 香芹酚可通过下调circ-0008717的表达而促进miR-217的表达从而减弱白血病细胞增殖、克隆形成能力及促进细胞凋亡.
Keywords: carvacrol; leukemia; cell apoptosis; cell proliferation; circ-0008717; miR-217.