基于Ti<sub>3</sub>C<sub>2</sub>负载阿霉素联合光热治疗抑制乳腺癌细胞增殖的研究

doi:10.3877/cma.j.issn.2095-5782.2023.02.008

目的

本研究基于Ti₃C₂二维纳米片装载阿霉素（doxorubicin，DOX）并包裹CaCO₃以制备新型复合纳米药物Ti₃C₂/DOX@CaCO₃，旨在增加药物在肿瘤部位的积聚，同时联合光热治疗增强乳腺癌的治疗效果，并减少DOX的毒副作用。

方法

以环己烷、Triton X-100、1-己醇、氯化钙溶液和碳酸钠溶液为原料制备Ti₃C₂/DOX@CaCO₃。采用CCK8实验检测Ti₃C₂/DOX@CaCO₃及联合光热治疗后对乳腺癌细胞MDA-MB-231增殖能力的影响。随后通过构建MDA-MB-231乳腺癌裸鼠皮下瘤模型，评估Ti₃C₂/DOX@CaCO₃联合光热治疗后对肿瘤生长的抑制效果以及生物安全性。

结果

Zeta电位显示Ti₃C₂成功负载DOX。CCK8实验结果表明，与单纯DOX处理组相比，Ti₃C₂/DOX@CaCO₃联合光热治疗后能更好地抑制乳腺癌细胞MDA-MB-231的增殖。同时体内实验证实，Ti₃C₂/DOX@CaCO₃联合光热治疗组相较单纯注射Ti₃C₂/DOX@CaCO₃组能更有效地抑制小鼠皮下肿瘤的生长，HE结果显示其他脏器未出现明显的形态改变和病理损伤。

结论

上述研究结果证实，通过Ti₃C₂二维纳米片递送化疗药物DOX同时联合光热治疗能够有效抑制乳腺癌细胞的生长，减少DOX对机体的毒副作用。这也为临床治疗乳腺癌提供了一种新型的治疗策略，在改善乳腺癌治疗疗效方面具有积极的临床应用前景。

关键词: 阿霉素, 碳化钛, 纳米药物, 乳腺癌细胞, 光热治疗

Objective

This study uses Ti₃C₂ nanosheets loading doxorubicin (DOX) and CaCO₃ to prepare a novel nano-composite Ti₃C₂/DOX@CaCO₃ for drug delivery, aiming to increase the accumulation of drugs at the site of tumors, enhance the therapeutic effect of breast cancer combined with photothermal therapy, and reduce the toxicity of DOX.

Methods

Ti₃C₂/DOX@CaCO₃ was prepared from Ti₃C₂, cyclohexane, Triton X-100, 1-hexanol, calcium chloride solution and sodium carbonate solution. CCK8 assay was used to detect the effect of Ti₃C₂/DOX@CaCO₃ combined with photothermal therapy on the proliferative ability of MDA-MB-231 cells. Subsequently, we established MDA-MB-231 tumor burden animal model. The toxicity of Ti₃C₂/DOX@CaCO₃ combined with photothermal therapy and the inhibitory effect of tumor growth in MDA-MB-231 breast cancer were evaluated by animal experiments.

Results

Zeta potential showed Ti₃C₂ loading DOX successfully.The CCK8 experiment confirmed that Ti₃C₂/DOX@CaCO₃ combined with photothermal therapy could better inhibit the proliferation of MDA-MB-231 cells compared with the DOX group. In vivo experiments, the results confirmed that Ti₃C₂/DOX@CaCO₃ nanocomposites combined with photothermal therapy could more effectively inhibit the growth of subcutaneous tumors in mice. Moreover, there were no other obvious morphological changes or pathological damage in other organs.

Conclusions

This study has confirmed that the Ti₃C₂ two-dimensional nanosheets loading chemotherapeutic drugs DOX and Ca²⁺, combined with photothermal therapy can effectively inhibit the growth of breast cancer cells, and reduce the toxicity of DOX. This also provides a new synergistic strategy for breast cancer treatment, which has a promising clinical application prospects.

Key words: Doxorubicin, Titanium carbide, Nanomedicine, Breast cancer, Photothermal therapy

图1 Ti₃C₂及Ti₃C₂/DOX@CaCO₃形貌及电位1A：Ti₃C₂透射电镜图像，比例尺：0.5 μm；1B：游离DOX、Ti₃C₂、Ti₃C₂/DOX及Ti₃C₂/DOX@CaCO₃的Zeta电位，n = 3,数据采用平均值±标准差。

图2 Ti₃C₂/DOX@CaCO₃联合光热治疗对MDA-MB-231细胞增殖活性的影响2A：特定浓度下MDA-MB-231细胞分别与游离DOX和Ti₃C₂/DOX@CaCO₃共同孵育24 h后的细胞活性情况；2B：特定浓度下MDA-MB-231细胞分别与游离DOX和Ti₃C₂/DOX@CaCO₃共同孵育48 h后的细胞活性情况；2C：MDA-MB-231细胞在不同治疗后细胞活性情况。n = 3，数据采用平均值±标准差；经one-way ANOVA分析，^***代表P < 0.001，^****表示P < 0.000 1。

图3 Ti₃C₂/DOX@CaCO₃及联合光热治疗对体内抗肿瘤效果分析3A：不同组别荷瘤老鼠治疗结束后肿瘤图片；3B：不同组别荷瘤老鼠的平均肿瘤生长曲线；3C：不同组别荷瘤老鼠结束治疗后的肿瘤重量对比柱形图；3D：不同组别荷瘤老鼠的平均体重变化曲线。n = 3，数据采用平均值±标准差，^*代表P < 0.05，^***代表P < 0.001。

图4 荷瘤老鼠治疗后各器官（心、肝、脾、肺、肾）HE染色

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