摘要
目的探讨石墨烯氧化物纳米颗粒负载NBDC6-神经酰胺(NPP/C)对乳腺癌细胞(Luminal A型)的抗肿瘤作用。方法制备NPP/C为实验组,以荧光标记的神经酰胺(NBDC6-Ceramide)为对照组,转染人乳腺癌细胞(MCF-7),荧光显微镜观察转运效率,细胞计数试剂盒(CCK-8)和克隆平板检测细胞增殖能力,Transwell迁移实验检测细胞移动能力,流式细胞术检测细胞凋亡,蛋白质印迹法(Western blot)检测相关凋亡蛋白,裸鼠皮下成瘤测量肿瘤增殖能力。组间比较采用t检验分析。结果NPP/C转运释放的神经酰胺效能明显高于NBD C6-Ceramide。CCK-8显示实验组和对照组细胞在不同时间吸光度(A)值分别为0.20±0.03比0.17±0.04(t=1.039,P>0.05),0.41±0.05比0.26±0.02(t=4.335,P<0.05),0.52±0.04比0.38±0.03(t=4.363,P<0.01)和1.25±0.06比0.59±0.07(t=8.128,P<0.01)。细胞克隆平板结果示两组细胞数分别为(577.3±41.0)个和(962.7±74.5)个(t=-7.850,P<0.01)。Transwell迁移实验提示实验组细胞移动能力明显低于对照组。流式细胞学检测两组细胞凋亡率分别为(79.29±8.44)%和(20.51±17.35)%(t=5.284,P<0.01)。Western blot检测实验组中凋亡相关蛋白Caspase-3和聚腺苷二磷酸核糖聚合酶(PARP)表达显著高于对照组,而B细胞淋巴瘤因子2(bcl-2)表达显著低于对照组(P<0.05)。裸鼠体内验证,两组皮下移植瘤体积分别为(942±73)mm^(3)和(1472±109)mm^(3)(t=8.265,P<0.01);实验组移植瘤中增殖细胞核抗原Ki-67表达显著低于对照组,而半胱氨酸蛋白酶-3(Caspase-3)表达高于对照组(P<0.05)。结论NPP/C纳米颗粒可抑制乳腺癌细胞(luminalA型)的增殖和迁移能力。
Objective To investigate the anti-tumor effect of nanoparticles graphene oxide-(polyethylene glycol)PEG-(polyethylenimine)PEI/ceramide(NPP/C)of luminal A breast cancer cells.Methods NPP/C(experimental group)and NBD C6-ceramide(control group)were synthesized and transfected into breast cancer cells(MCF-7).In vitro,the transport efficiency was observed by fluorescence microscopy,cell counting kit(CCK-8)and clone plate assay were applied to measure cell proliferation,Transwell was used to detect the mobility of cells,flow cytometry was employed to test cell apoptosis,and Western blotting was utilized to detect related apoptotic proteins.In vivo,subcutaneous xenografts in nude mice were utilized to detect tumor proliferation.Comparisons between groups were performed by t test.Results The transfection efficiency of NPP/C was significantly higher than NBD C6-ceramide.Absorbance(A)values of the two groups detected by CCK-8 at 24,48,72,96 h were 0.20±0.03 vs.0.17±0.04(t=1.039,P>0.05),0.41±0.05 vs.0.26±0.02(t=4.335,P<0.05),0.52±0.04 vs.0.38±0.03(t=4.363,P<0.01)and 1.25±0.06 vs.0.59±0.07(t=8.128,P<0.01),respectively.Cell counts were(577.3±41.0)vs.(962.7±74.5)in two groups(t=-7.850,P<0.01).The mobility of cells in experimental group was significantly reduced as compared with that in control group.The apoptosis rates in two groups were(79.29±8.44)%vs.(20.51±17.35)%(t=5.284,P<0.01),respectively.The expression levels of Caspase-3 and poly ADP ribose polymerase(PARP)were significantly increased,and those of B-cell lymphoma-2(bcl-2)significantly decreased in experimental group.In vivo,the volume of xenografts in two groups were(942±73)mm3 vs.(1472±109)mm3(t=8.265,P<0.01).The expression of Ki-67 was down-regulatied in experimental group.On the contrary,the expression of Caspase-3 was up-regulation(P<0.05).Conclusion NPP/C can play a tumor suppressive role by inhibiting the proliferation and migration of luminal A breast cancer cells.
作者
李永强
刘小平
曾维乾
陈友科
林大宁
杨彦
Li Yongqiang;Liu Xiaoping;Zeng Weiqian;Chen Youke;Lin Daning;Yang Yan(Department of Hepatobiliary Pancreatic Surgery,the First Affiliated Hospital of Medical University,Haikou 570102,China;Department of Breast Surgery,Haikou People’s Hospital,Haikou 570208,China)
出处
《中华实验外科杂志》
CAS
北大核心
2022年第4期629-632,共4页
Chinese Journal of Experimental Surgery
基金
国家自然科学基金地区基金项目(81860514)
海南省自然科学基金高层次人才项目(2019RC380)。
