Pineda-Castellanos Mónica L, Santa-Olalla-Tapia Jesús, de C Gutiérrez María, González-Maya Leticia, Sánchez-Carranza Jessica N, Nuñez-Valdez M Eugenia
Laboratorio de Patogenicidad Molecular, Centro de Investigación en Dinámica Celular, Universidad Autónoma del Estado de Morelos, Av. Universidad 1001, Col. Chamilpa, 62209, Cuernavaca, Morelos, México.
Laboratorio de Biología de Células Troncales, Facultad de Medicina, Universidad Autónoma del Estado de Morelos, Cuernavaca, Morelos, México.
Arch Microbiol. 2023 Mar 3;205(4):103. doi: 10.1007/s00203-023-03443-w.
It has been reported that cell-free culture broths and some proteins from pigmented and non-pigmented Serratia spp. are cytotoxic towards cancerous and non-cancerous human cell lines. Looking for new molecules toxic against human cancerous cells but harmless towards normal human cells, the aim of this work was (a) to determine whether cell-free broths from the entomopathogenic non-pigmented S. marcescens 81 (Sm81), S. marcescens 89 (Sm89) and S. entomophila (SeMor4.1) presented cytotoxic activity towards human carcinoma cell lines; (b) to identify and purify the associated cytotoxic factor(s) and (c) to evaluate whether the cytotoxic factor(s) was cytotoxic towards non-cancerous human cells. This research was focussed on the observed morphology changes and the proportion of remaining viable cells after incubation in the presence of cell-free culture broths from the Serratia spp isolates to evaluate cytotoxic activity. The results showed that broths from both S. marcescens isolates presented cytotoxic activity and induced cytopathic-like effects on the human neuroblastoma CHP-212 and the breast cancer MDA-MB-231 cells. Slight cytotoxicity was observed in the SeMor4.1 broth. A serralysin-like protein of 50 kDa was identified in Sm81 broth as responsible for cytotoxic activity after purification by ammonium sulphate precipitation and ion-exchange chromatography followed by tandem-mass spectrometry (LC-MS/MS). The serralysin-like protein was toxic against CHP-212 (neuroblastoma), SiHa (human cervical carcinoma) and D-54 (human glioblastoma) cell lines in a dose-dependent manner and showed no cytotoxic activity in primary cultures of normal non-cancerous human keratinocytes and fibroblasts. Therefore, this protein should be evaluated for a potential use as an anticancer agent.
据报道，来自产色素和不产色素的沙雷氏菌属的无细胞培养液和一些蛋白质对人癌细胞系和非癌细胞系具有细胞毒性。为了寻找对人癌细胞有毒但对正常人细胞无害的新分子，本研究的目的是：（a）确定来自昆虫致病性无色素粘质沙雷氏菌81（Sm81）、粘质沙雷氏菌89（Sm89）和嗜虫沙雷氏菌（SeMor4.1）的无细胞培养液是否对人癌细胞系具有细胞毒性活性；（b）鉴定和纯化相关的细胞毒性因子；（c）评估细胞毒性因子对非癌人细胞是否具有细胞毒性。本研究聚焦于在存在沙雷氏菌属分离株的无细胞培养液孵育后观察到的形态变化和剩余活细胞的比例，以评估细胞毒性活性。结果表明，两种粘质沙雷氏菌分离株的培养液均表现出细胞毒性活性，并对人神经母细胞瘤CHP - 212和乳腺癌MDA - MB - 231细胞诱导了细胞病变样效应。在SeMor4.1培养液中观察到轻微的细胞毒性。通过硫酸铵沉淀和离子交换色谱法，随后进行串联质谱（LC - MS/MS）纯化后，在Sm81培养液中鉴定出一种50 kDa的类丝氨酸蛋白酶，其负责细胞毒性活性。该类丝氨酸蛋白酶对CHP - 212（神经母细胞瘤）、SiHa（人宫颈癌）和D - 54（人胶质母细胞瘤）细胞系具有剂量依赖性毒性，并且在正常非癌人角质形成细胞和成纤维细胞的原代培养中未显示细胞毒性活性。因此，该蛋白应评估其作为抗癌剂的潜在用途。
