Data_Sheet_1_Comparative Transcriptomics Unravels Prodigiosin's Potential Cancer-Specific Activity Between Human Small Airway Epithelial Cells and Lung Adenocarcinoma Cells.docx

Objective: Non-Small Cell Lung Cancer (NSCLC) is extremely lethal upon metastasis and requires safe and effective systemic therapies to improve a patient's prognosis. Prodigiosin (PG) appears to selectively and effectively target cancer but not healthy cells. However, PG's cancer-specific activity has remained elusive until recently.
Methods: PG's cancer-specific performance was compared to Docetaxel (DTX), Paclitaxel (PTX), and Doxorubicin (DOX) against human lung adenocarcinoma (A549) and human small airway epithelial cells (HSAEC). Combination of PG with DTX, PTX, or DOX in a 1:1 ED50 ratio was also evaluated. MTT assay was used to determine the post-treatment cell viability. RNA-sequencing was used for comparative transcriptomics analysis between A549 and HSAEC treated with 1.0 μM PG for 24 h.
Results: PG reduced A549 cell viability by four-folds greater than HSAEC. In comparison to DTX, PTX and DOX, PG was ~1.7 times more toxic toward A549, and 2.5 times more protective toward HSAEC. Combination of PG in a 1:1 ED50 ratio with DTX, PTX, or DOX failed to exhibit synergistic toxicity toward A549 or protection toward HSAEC. In A549, genes associated in DNA replication were downregulated, while genes directly or indirectly associated in lipid and cholesterol biogenesis were upregulated. In HSAEC, co-upregulation of oncogenic and tumor-suppressive genes was observed.
Conclusion: An overactive lipid and cholesterol biogenesis could have caused A549's autophagy, while a balancing-act between genes of oncogenic and tumor-suppressive nature could have conferred HSAEC heightened survival. Overall, PG appears to be a smart chemotherapeutic agent that may be both safe and effective for NSCLC patients.

研究目的：非小细胞肺癌（Non-Small Cell Lung Cancer, NSCLC）发生转移后致死性极强，亟需安全有效的全身治疗方案以改善患者预后。灵菌红素（Prodigiosin, PG）可选择性且高效地靶向杀伤癌细胞而不损伤健康细胞，但直至近期，其癌症特异性活性的分子机制仍未阐明。
研究方法：本研究将灵菌红素（PG）与多西他赛（Docetaxel, DTX）、紫杉醇（Paclitaxel, PTX）及多柔比星（Doxorubicin, DOX）进行对比，评估其对人肺腺癌细胞（A549）与人小气道上皮细胞（HSAEC）的作用效果；同时以1:1的半数有效剂量（ED50）比例，将PG分别与DTX、PTX及DOX联合给药，评估联合给药的效应。采用MTT法检测给药后细胞活力；以1.0 μM浓度的PG处理A549与HSAEC细胞24小时后，通过RNA测序（RNA-sequencing）开展比较转录组学分析。
研究结果：PG对A549细胞活力的抑制效果较HSAEC高4倍。与DTX、PTX及DOX相比，PG对A549的细胞毒性高约1.7倍，而对HSAEC的细胞保护作用高2.5倍。以1:1 ED50比例将PG与DTX、PTX或DOX联合给药时，未对A549表现出协同细胞毒性，亦未增强对HSAEC的保护作用。在A549细胞中，与DNA复制相关的基因表达下调，而直接或间接参与脂质与胆固醇生物合成的基因表达上调。在HSAEC细胞中，可观察到致癌基因与抑癌基因的共同上调现象。
研究结论：A549细胞的自噬可能由过度激活的脂质与胆固醇生物合成通路所介导，而HSAEC细胞中致癌基因与抑癌基因的平衡调控，可能赋予其更强的存活能力。综上，灵菌红素（PG）是一种兼具安全性与有效性的智能化疗药物，有望为非小细胞肺癌患者提供新的治疗选择。