SWATH-MS proteomics of PANC-1 and MIA PaCa-2 pancreatic cancer cells allows identification of drug targets alternative to MEK and PI3K inhibition.

Pancreatic cancer remains one of the most lethal diseases with dismal five-year survival rates. Although mutant KRas protein-driven activation of downstream MAPK Raf/MEK/ERK and PI3K/Akt signaling pathways represent major oncogenic alterations, signaling blockade with MEK and PI3K inhibitors has shown that intrinsic resistance may hamper the effectiveness of this targeted approach. However, there have been no mass spectrometry-based proteomic studies for in-depth comparison of protein expression differences between pancreatic cancer cells with sensitivity and resistance to MEK and PI3K kinase inhibitors. In this work, we compared PANC-1 and MIA PaCa-2 pancreatic cancer cells which are, respectively, resistant and sensitive to MEK- and PI3K-targeted therapy. We conducted a label-free data-independent acquisition mass spectrometry (SWATH-MS) study with extensive peptide fractionation to quantitate 4808 proteins and analyze differential expression of 743 proteins between resistant and sensitive cells. This allowed identification of the tumor suppressor protein phosphatase 2A (PP2A) and proteins from mitochondrial respiratory complex I implicated in oxidative phosphorylation as alternative candidate drug targets for cells resistant to MEK and PI3K inhibition. PP2A activator DT-061 decreased viability of PANC-1 cells and this was accompanied by reduced expression of c-Myc. PANC-1 cells also showed response to metformin and the novel complex I inhibitor IACS-010759. These findings provide insights into the distinct cellular proteomes and point out alternative pharmacological targets for MEK and PI3K inhibition-resistant pancreatic cancer cells.

DERIVING PRIMARY CANCER CELL CULTURES FOR PERSONALIZED THERAPY.

Cancer is the second-leading cause of death in the world, accounting for one out of six deaths. Consequently, there is an urgent need for new and more effective therapeutic options as well as drug screening methods. Immortal, "stable" cancer cell lines have been employed since the past century to assess drug response but face several disadvantages. They often accumulate new genetic aberrations due to long-term culture and lack the indisputable heterogeneity of solid tumors, therefore, compromising the recapitulation of molecular features from parental tumors. Primary cancer cells have emerged as an attractive alternative to commercial cell lines since they can preserve such properties more closely. Here, we provide an overview of the basic concepts underlying generation and characterization of primary cell cultures from tumor samples. We emphasize the advantages and disadvantages of using these types of cancer cell cultures, and we make a comparison with other types of cultures used for personalized therapy. Finally, we consider the use of primary cancer cell cultures in personalized therapy as a means to improve drug response prediction and therapeutic outcomes.

Deriving primary cancer cell cultures for personalized therapy

ABSTRACT Cancer is the second-leading cause of death in the world, accounting for one out of six deaths. Consequently, there is an urgent need for new and more effective therapeutic options as well as drug screening methods. Immortal, “stable” cancer cell lines have been employed since the past century to assess drug response but face several disadvantages. They often accumulate new genetic aberrations due to long-term culture and lack the indisputable heterogeneity of solid tumors, therefore, compromising the recapitulation of molecular features from parental tumors. Primary cancer cells have emerged as an attractive alternative to commercial cell lines since they can preserve such properties more closely. Here, we provide an overview of the basic concepts underlying generation and characterization of primary cell cultures from tumor samples. We emphasize the advantages and disadvantages of using these types of cancer cell cultures, and we make a comparison with other types of cultures used for personalized therapy. Finally, we consider the use of primary cancer cell cultures in personalized therapy as a means to improve drug response prediction and therapeutic outcomes.

Establecimiento y caracterización de la línea celular CaLo y del CLON Kalo, obtenidos a partir de un carcinoma de cervix, y efecto de IL-2, IL-3, GM-CSF, M-CSF, TNF-Ó E IFN-ç sobre su proliferación / Establishing and characterization of the CaLo cell line and KaLo clon, ontained from a cervix carcinoma and IL-2, IL-3, GM-CSF, M-CSF, TNF-Ó and IFN-ç on it's proliferation

En este trabajo se describe la caracterización morfológica, cromosómica y presencia de desmosomas de una línea celular (CaLo) derivada de una biopsia de cáncer cervicouterino. Nuestros resultados indican que CaLo posee una morfología típica de células epiteliales, una aneuploidia cromosómica con número modal de 58 y presencia de desmosomas de bida a la positividad en membrana para la presencia de desmogleína-1. A partir de CaLo también se aisló un clon llamado KaLo. esta clonación nos proporciona una evidencia del origen maligno de estas células. Ambos tipos celulares fueron cultivados en presencia de algunas citocinas recientemente empleadas en ciertos protocolos clínicos (TNF-Ó, IL-2. IL-3 GM-CSF, M-CSF e IFN-ç). Nuestros resultados demuestran un efecto inhibidor de la proliferación por parte de TNF-Ó, IL-3 y GM-CSF, mientras que con M-CSF e IFN-ç no se detectó efecto. Por otra parte, obresvamos un incremento en la proliferación celular en presencia de la IL-2. Estos resultados dan indicios de que el TNF-Ó, la IL-3 y el M-CSF pueden tener posibilidades de aplicación clínica por sus propiedades inhibidoras; mientras que ponen en evidencia el riesgo de utilizar in vivo la IL-2, pues activa la proliferación de células tumorales de cérvix, tal como se ha informado para algunos carcinomas de cabeza y cuello