新型纳米材料石墨烯量子点对膀胱癌T24细胞作用的研究

EFFECT OF THE NOVEL NANOMATERIAL GRAPHENE QUANTUM DOTS ON BLADDER CANCER T24 CELLS

目的观察新型纳米材料石墨烯量子点(GQD)对膀胱癌T24细胞的杀伤作用,并探讨其作用机制。方法将T24细胞培养至对数生长期以后,细胞分为6组,分别为细胞对照组(A组)、20 mg/L GQD对照组(B组)、1 J/cm^2激光对照组(C组)、5 mg/L GQD+1 J/cm^2激光处理组(D组)、10 mg/L GQD+1 J/cm^2激光处理组(E组)、20 mg/L GQD+1 J/cm^2激光处理组(F组)。B、D、E、F组加入相应浓度GQD共同孵育2 h后,C、D、E、F组使用532 nm波长、能量密度为1 J/cm^2的激光照射后再培养12 h,采用CCK-8法检测GQD对T24细胞生长的抑制情况。将培养至对数生长期T24细胞加入10 mg/L GQD共同孵育2 h后,再加入DAPI进行T24细胞染色,激光共聚焦显微镜下双通道均选择波长405 nm激光激发,观察GQD在T24细胞内的定位情况。结果各组吸光度值比较差异有显著性(F=30.060,P<0.05),其中A、B、C、D组间比较,差异无显著性(P>0.05),E组与A、B、C、D组间比较,差异均有显著性(P<0.05),F组与A、B、C、D、E组间比较,差异均有显著性(P<0.05)。激光共聚焦显微镜观察发现,GQD发出的绿色荧光与DAPI发出的蓝色荧光位置基本重合。结论光动力作用下新型纳米材料石墨烯量子点对膀胱癌T24细胞有明确的杀伤作用,其进入细胞后主要定位并作用于细胞核。

Objective To investigate the killing effect of the novel nanomaterial graphene quantum dots(GQD)on bladder cancer T24 cells and the potential mechanism.Methods T24 cells were cultured to the logarithmic growth phase and then they were divided into cell control group(group A),20 mg/L GQD control group(group B),1 J/cm^2 laser control group(group C),5 mg/L GQD+1 J/cm^2 laser treatment group(group D),10 mg/L GQD+1 J/cm^2 laser treatment group(group E),and 20 mg/L GQD+1 J/cm^2 laser treatment group(group F).T24 cells in groups B,D,E,and F were incubated with GQD at the corresponding concentration for 2 h,and then T24 cells in groups C,D,E,and F were irradiated by laser at a wavelength of 532 nm and a energy density of 1 J/cm^2 and were further incubated for 12 h;CCK-8 assay was used to observe the inhibitory effect of GQD on the growth of T24 cells.T24 cells in the logarithmic growth phase were incubated with 10 mg/L GQD for 2 h and then stained with DAPI,and T24 cells were excited with the 405 nm dual-channel laser to observe the localization of GQD in cells under a laser scanning confocal microscope.Results There was a significant difference in absorbance between groups(F=30.060,P<0.05),with no significant difference between groups A,B,C,and D(P>0.05),and there was a significant difference between group E and groups A,B,C,and D(P<0.05)and between group F and groups A,B,C,D,and E(P<0.05).The green fluorescence emitted by GQD and the blue fluorescence emitted by DAPI had almost the same location under the laser scanning confocal microscope.Conclusion The novel nanomaterial GQD has a marked killing effect on bladder cancer T24 cells under the photo-dynamic effect,and GQD is mainly localized in the nucleus and acts on it after entering the cell.