HC-NPs对RAW264.7-4T1共培养体系中乳腺癌细胞增殖及凋亡的影响

Effects of Hydrazinocurcumin-loaded Nanoparticles on the Proliferation and Apoptosis of Breast Cancer Cells in RAW264.7- 4T1 Coculture Conditions

目的:探索联氨基姜黄素纳米脂质体(hydrazinocurcumin-loaded nanopaticles,HC-NPs)通过抑制STAT3(signal trannsduction and activators oftranscription-3)活化,对RAW264.7-4T1共培养体系中4T1细胞增殖及凋亡的影响。方法:以乳腺癌细胞—巨噬细胞共培养体系中的乳腺癌细胞4T1为研究对象,分PBS对照组和10%NPs对照组、10%HC-NPs处理组采用检测细胞形态学变化,台盼蓝染色计数细胞存活率,流式细胞仪检测细胞凋亡率及细胞周期变化,Western blot检测STAT3、p-STAT3、c-MYC、Bcl-2及Bax的蛋白表达水平。结果:刘氏染色发现经10%HC-NPs处理后,4T1细胞形态变圆,台盼蓝染色计数显示细胞存活率减低,且与对照组有显著性差异(P<0.05);流式细胞仪检测发现HC-NPs处理组细胞凋亡率为(24.21±2.37)%,细胞周期结果显示HC-NPs组处理的细胞主要阻滞在G_2/M期为(63.70±2.53)%,与对照组有显著性差异(P<0.05);Western blot检测显示,HC-NPs组4T1细胞的p-STAT3、c-MYC表达明显减少,Bcl-2/Bax比值下降,而总STAT3水平无明显变化。结论:HC-NP_s能通过抑制STAT3活化(p-STAF3),影响细胞增殖、凋亡相关基因,抑制共培养体系中乳腺癌细胞4T1的增殖,促进其凋亡。

Objective： To investigate the effects of hydrazinocurcumin-loaded nanoparticles （ HC-NPs ） on the proliferation and apoptosis of breast cancer 4T 1 cells in coculture conditions through the inhibition of the signal transducer and activator of transcription 3 （ STAT3 ） signal pathway activation. Methods： Mouse breast cancer 4T1 cells in coculture conditions were divided into three groups, namely, the PBS control, 10% NP control, and 10% HC-NP treatment groups. Morphological investigation was conducted via Liu staining, and the survival ratio was determined via trypan blue staining. Cell cycle and apoptosis were detected using flow cytometry. Western blot analysis was used to determine the expression of STAT3, phosphorylation STAT3 （ pSTAT3 ）, c-MYC, B-cell lymphoma 2 （ Bcl-2 ）, and Bcl-2-associated X protein （ Bax ）. Results： Liu staining results show that the shape of the HC-NP-treated 4T1 ceils was spherical. Moreover, trypan blue staining shows that the survival ratio of the treated cells was significantly decreased compared with that of the control. Flow cytometry results show that the apoptosis rates of the HC-NP treatment groups was 24.21%±2.37% （ P 〈 0.05 ）, whereas that of The cells in the GIM phase increased （ 63.70%±2.53%, P 〈 0.05 ）. Western blot analysis indicates that p-STAT3, c-MYC, and the ratio of Bcl-2/Bax expression decreased. However, that of total STAT3 showed no significant difference with that of the control. Conclusion： HC-NPs can inhibit the proliferation and promote the apoptosis of breast cancer 4T1 cells in coculture conditions, possibly by inhibiting STAT3 activity and affecting the genes related to proliferation and apoptosis.