Showa University School of Medicine

3 ARTICLES PUBLISHED IN JoVE

Cancer Research

Establishing Dual Resistance to EGFR-TKI and MET-TKI in Lung Adenocarcinoma Cells In Vitro with a 2-step Dose-escalation Procedure

Toshimitsu Yamaoka¹, Motoi Ohba¹, Satoru Arata^1,2, Tohru Ohmori¹

¹Institute of Molecular Oncology, Showa University, ²Center for Biotechnology, Showa University

An in vitro method for establishing dual resistance to an EGFR-TKI and a MET-TKI in cancer cells is described. This method is useful for developing treatments for patients with EGFR-mutations, who exhibit disease progression despite EGFR-TKI treatment with MET-amplification. It can also be modified for inhibitors targeting other molecules.

Cancer Research

Establishment and Characterization of Three Afatinib-resistant Lung Adenocarcinoma PC-9 Cell Lines Developed with Increasing Doses of Afatinib

Toshimitsu Yamaoka¹, Motoi Ohba¹, Yuki Matsunaga², Junji Tsurutani¹, Tohru Ohmori³

¹Advanced Cancer Translational Research Institute, Showa University, ²Division of Breast Surgical Oncology, Department of Surgery, Showa University School of Medicine, ³Division of Allergology and Respiratory Medicine, Department of Medicine, Showa University School of Medicine

A method for establishing afatinib-resistance cell lines from lung adenocarcinoma PC-9 cells was developed, and resistant cells were characterized. The resistant cells can be used to investigate epidermal growth factor receptor tyrosine kinase inhibitor-resistance mechanisms, applicable for patients with non-small cell lung cancer.

JoVE Journal

Evaluation of Protein–Protein Interactions using an On-Membrane Digestion Technique

Takashi Obama^*1, Takuro Miyazaki^*2, Toshihiro Aiuchi¹, Akira Miyazaki², Hiroyuki Itabe¹

¹Division of Biological Chemistry, Department of Molecular Biology, Showa University School of Pharmacy, ²Department of Biochemistry, Showa University School of Medicine

Here, we present a protocol for an on-membrane digestion technique for the preparation of samples for mass spectrometry. This technique facilitates the convenient analysis of protein–protein interactions.