TMED3 promotes the progression and development of lung squamous cell carcinoma by regulating EZR

An Xie,Ling Zhang,Xinping Xu,Peng Kuang,Feng Yu

doi:10.1038/s41419-021-04086-9

Abstract

Lung squamous cell carcinoma (LUSC) has a poor clinical prognosis and lacks effective targeted therapy. The transmembrane emp24 trafficking protein 3 (TMED3) belongs to the TMED family, which is responsible for the transport of intracellular proteins. This study was to explore the clinicopathological significance and biological effects of TMED3 in LUSC. Expression of TMED3 in LUSC was detected by immunohistochemical (IHC). The loss-of-function assays were used to investigate the effects of TMED3 on proliferation, apoptosis, cell cycle, and migration of LUSC cells. The influence of TMED3 knockdown on tumor growth in vivo was evaluated by mice xenograft models. In addition, the downstream target of TMED3 was recognized by RNA sequencing and Ingenuity Pathway Analysis (IPA). Moreover, TMED3 was upregulated in LUSC tissue, which was positively correlated with pathological grade. TMED3 knockdown was involved in the regulation of LUSC cell function, such as inhibition of proliferation, reduction of colony formation, induction of apoptosis, and reduction of migration. TMED3 knockdown induced abnormalities in apoptosis-related proteins in LUSC cells. In addition, the inhibition of cell migration by TMED3 knockdown was achieved by regulating EMT. Mechanically, EZR was considered as a potential target for TMED3 to regulate the progress of LUSC. Inhibition of EZR can inhibit the progression of LUSC, and even reduce the promoting effects of TMED3 overexpression on LUSC. In conclusion, TMED3 promoted the progression and development of LUSC by EZR, which may be a novel therapeutic target for LUSC.

Highlights

Lung cancer has become the leading cause of death, with more deaths than colorectal, breast, prostate, and pancreatic cancer combined, and nearly 1.8 million people are diagnosed with lung cancer each year [1]
In order to further clarify the role of TMED3 in Lung squamous cell carcinoma (LUSC), we first detected the expression of TMED3 in clinical LUSC patients by immunohistochemistry
The results in vivo demonstrated that TMED3 knockdown had an inhibitory effect on the proliferation of cancer cells, these results indicate the promotive role of TMED3 in LUSC

Summary

INTRODUCTION

Lung cancer has become the leading cause of death, with more deaths than colorectal, breast, prostate, and pancreatic cancer combined, and nearly 1.8 million people are diagnosed with lung cancer each year [1]. Transwell invasion assay According to the method provided in the literature [20], the suspension of cells (EBC-1, NCI-H520, and SK-MES-1) infected with TMED3 (overexpression), shEZR, TMED3+shEZR, and the corresponding negative controls were first adjusted to 5 × 105 cells/mL, and the cells were seeded into Transwell (Corning, NY, USA). Quantitative real time PCR (qPCR) Total RNA was extracted from the EBC-1, NCI-H520 and SK-MES-1 cells infected with TMED3 (over-expression), shEZR, TMED3+shEZR, and the corresponding negative controls using TRIzol® Western blot analysis Protein was extracted from the cells EBC-1, NCI-H520, and SK-MES-1 infected with TMED3 over-expression, shEZR, TMED3+shEZR and the corresponding negative controls) using lysis buffer (Nanjing KeyGen Biotech, Nanjing, Jiangsu, China), and protein concentration is measured using the BCA protein assay kit (Takara, Otsu, Japan).

Xie et al 3

RESULTS

DISCUSSION

10 COMPETING INTERESTS