Gatti Laura, Rolli Luigi, Corno Cristina, Carenini Nives, Corna Elisabetta, Ciusani Emilio, Frigerio Simona, Pogliani Simona, Guarino Carmela, Ravagnani Ferdinando, Pastorino Ugo, Sozzi Gabriella, Macciotta Alessandra, Verderio Paolo, Ciniselli Chiara M, Perego Paola
Molecular Pharmacology Unit, Fondazione IRCCS Istituto Nazionale dei Tumori, Milan, Italy.
Neurobiology Laboratory, Neurology IX, Fondazione IRCCS Istituto Neurologico Carlo Besta, Milan, Italy.
Transl Lung Cancer Res. 2022 Jul;11(7):1315-1326. doi: 10.21037/tlcr-22-52.
The secreted products of the metastasis suppressor gene may represent useful biomarkers in non-small cell lung cancer (NSCLC) but their levels in patients have remained poorly investigated. We previously found that forced expression of decreased the invasive capability of NSCLC drug-resistant cells and a pro-apoptotic role for KiSS1 has been proposed in head and neck cancer. Thus, we designed a translational investigation including a pilot study to analyze KiSS1 levels in liquid biopsies, and experiments to explore the biological relevance of KiSS1 modulation.
KiSS1-derived peptide levels in liquid biopsies from 60 NSCLC patients were assayed by ELISA. Preclinical experiments were carried out using quantitative real time polymerase chain reaction (qRT-PCR), ELISA, annexin V-binding and caspase activation assays.
We compared KiSS1 release in 3 different matrices (serum, plasma and urine) and the highest levels were detectable in serum (range, 0-4.5 ng/mL). We observed increased levels of seric KiSS1 in NSCLC patients as compared to healthy donors. KiSS1 serum concentrations, after surgical procedure and/or adjuvant therapy. We observed differences among disease stages in urine samples. In preclinical models, mRNA levels were increased by short term exposure to azacytidine, enhanced KiSS1 release was induced by the combination of azacytidine and cisplatin and KiSS1-derived peptides enhanced cisplatin-induced apoptosis. KiSS1 increase was observed upon exposure neurons-enriched cultures to tumor cell conditioned medium.
Our results showing a peculiar modulation of KiSS1 levels in liquid biopsies of NSCLC patients and a regulation of cisplatin-induced apoptosis by KiSS1-derived peptides support an involvement of KiSS1 in cell response to treatment and highlight its promising features as a potential biomarker in NSCLC.
转移抑制基因的分泌产物可能是非小细胞肺癌（NSCLC）中有用的生物标志物，但其在患者体内的水平仍研究不足。我们之前发现，强制表达可降低NSCLC耐药细胞的侵袭能力，并且有人提出KiSS1在头颈癌中具有促凋亡作用。因此，我们设计了一项转化研究，包括一项初步研究以分析液体活检中的KiSS1水平，以及实验以探索KiSS1调节的生物学相关性。
采用酶联免疫吸附测定（ELISA）法检测60例NSCLC患者液体活检中KiSS1衍生肽的水平。使用定量实时聚合酶链反应（qRT-PCR）、ELISA、膜联蛋白V结合和半胱天冬酶激活测定进行临床前实验。
我们比较了3种不同基质（血清、血浆和尿液）中KiSS1的释放情况，血清中可检测到的水平最高（范围为0 - 4.5 ng/mL）。与健康供体相比，我们观察到NSCLC患者血清中KiSS1水平升高。手术后和/或辅助治疗后KiSS1血清浓度。我们观察到尿液样本中疾病分期之间存在差异。在临床前模型中，短期暴露于阿扎胞苷可使mRNA水平升高，阿扎胞苷和顺铂联合使用可诱导KiSS1释放增强，并且KiSS1衍生肽可增强顺铂诱导的细胞凋亡。将富含神经元的培养物暴露于肿瘤细胞条件培养基后观察到KiSS1增加。
我们的结果显示NSCLC患者液体活检中KiSS1水平有特殊调节，并且KiSS1衍生肽可调节顺铂诱导的细胞凋亡，这支持KiSS1参与细胞对治疗的反应，并突出了其作为NSCLC潜在生物标志物的前景。
