Qi Yuxin, Yang Wenping, Liu Shuang, Han Fanjie, Wang Haibin, Zhao Yonghong, Zhou Yufa, Zhou Daijun
Department of Respiratory Medicine, Jinan People's Hospital Affiliated to Shandong First Medical University, Jinan, 271199, China.
Department of Oncology, General Hospital of Western Theater Command of PLA, Chengdu, 610083, China.
Cancer Cell Int. 2021 Jan 25;21(1):74. doi: 10.1186/s12935-021-01771-9.
Lung cancer is one of the important health threats worldwide, of which 5-year survival rate is less than 15%. Non-small-cell lung cancer (NSCLC) accounts for about 80% of all lung cancer with high metastasis and mortality.
Cisplatin loaded multiwalled carbon nanotubes (Pt-MWNTS) were synthesized and used to evaluate the anticancer effect in our study. The NSCLC cell lines A549 (cisplatin sensitive) and A549/DDP (cisplatin resistant) were used in our in vitro assays. MTT was used to determine Cancer cells viability and invasion were measured by MTT assay and Transwell assay, respectively. Apoptosis and epithelial-mesenchymal transition related marker proteins were measured by western blot. The in vivo anti-cancer effect of Pt-MWNTs were performed in male BALB/c nude mice (4-week old).
Pt-MWNTS were synthesized and characterized by X-ray diffraction, Raman, FT-IR spectroscopy and scan electron microscopy. No significant cytotoxicity of MWNTS was detected in both A549/DDP and A549 cell lines. However, Pt-MWNTS showed a stronger inhibition effect on cell growth than free cisplatin, especially on A549/DDP. We found Pt-MWNTS showed higher intracellular accumulation of cisplatin in A549/DDP cells than free cisplatin and resulted in enhanced the percent of apoptotic cells. Western blot showed that application of Pt-MWNTS can significantly upregulate the expression level of Bax, Bim, Bid, Caspase-3 and Caspase-9 while downregulate the expression level of Bcl-2, compared with free cisplatin. Moreover, the expression level of mesenchymal markers like Vimentin and N-cadherin was more efficiently reduced by Pt-MWNTS treatment in A549/DDP cells than free cisplatin. In vivo study in nude mice proved that Pt-MWNTS more effectively inhibited tumorigenesis compared with cisplatin, although both of them had no significant effect on body weight.
Pt-MWNT reverses the drug resistance in the A549/DDP cell line, underlying its possibility of treating NSCLC with cisplatin resistance.
肺癌是全球重要的健康威胁之一，其5年生存率低于15%。非小细胞肺癌（NSCLC）约占所有肺癌的80%，具有高转移率和高死亡率。
在本研究中合成了负载顺铂的多壁碳纳米管（Pt-MWNTS）并用于评估其抗癌效果。体外实验使用了NSCLC细胞系A549（顺铂敏感型）和A549/DDP（顺铂耐药型）。采用MTT法测定癌细胞活力，分别通过MTT法和Transwell法检测侵袭能力。通过蛋白质免疫印迹法检测凋亡及上皮-间质转化相关标记蛋白。在4周龄雄性BALB/c裸鼠体内进行Pt-MWNTs的抗癌效果实验。
通过X射线衍射、拉曼光谱、傅里叶变换红外光谱和扫描电子显微镜对合成的Pt-MWNTS进行了表征。在A549/DDP和A549细胞系中均未检测到MWNTS有明显的细胞毒性。然而，Pt-MWNTS对细胞生长的抑制作用比游离顺铂更强，尤其是对A549/DDP细胞。我们发现Pt-MWNTS在A549/DDP细胞中顺铂的细胞内积累量比游离顺铂更高，并导致凋亡细胞百分比增加。蛋白质免疫印迹显示，与游离顺铂相比，应用Pt-MWNTS可显著上调Bax、Bim、Bid、Caspase-3和Caspase-9的表达水平，同时下调Bcl-2的表达水平。此外，在A549/DDP细胞中，Pt-MWNTS处理比游离顺铂更有效地降低了波形蛋白和N-钙黏蛋白等间质标记物的表达水平。裸鼠体内研究证明，与顺铂相比，Pt-MWNTS能更有效地抑制肿瘤发生，尽管它们对体重均无显著影响。
Pt-MWNT可逆转A549/DDP细胞系中的耐药性，这表明其具有治疗顺铂耐药NSCLC的可能性。
