Peptide 17, an inhibitor of YAP/TEAD4 pathway, mitigates lung cancer malignancy

Jirong Zhang,Dehua Liao,Jingyi Tang,Dunwu Yao,Yong Pan

doi:10.4314/tjpr.v17i7.5

Abstract

Purpose: To investigate whether and how peptide 17 affects lung cancer cells.Methods: Human lung carcinoma cells, LLC and PC-9, were employed to study the therapeutic effect of peptide 17 on lung cancer. After exogenous expression of peptide 17, a co-immunoprecipitation experiment was used to examine the inhibitory effect of peptide 17. CCK8 assay was employed to assess the lung cancer cells’ viability while clone formation assays were used to assess lung cancer cell proliferation. Colony number was also determined. The stimulatory effect of peptide 17 on lung cancer cell apoptosis was assessed by fluorescence-activated cell sorting (FACS).Results: Peptide 17 efficiently disrupted the interaction between YAP and TEAD4 (p < 0.001), and decreased the expression of CTGF and Cyr61. In addition, lung cancer cell viability and proliferation significantly decreased (p < 0.001) in a time- and concentration-dependent manner. On the other hand, the proportion of apoptotic cells was significantly elevated with rising concentration of peptide 17.Conclusion: Exogenous expression of peptide 17 activates Bcl2/Bax/caspase-9 signal and isresponsible for its inhibitory effects on lung cancer cells. Thus, peptide 17 is a promising target drug in lung cancer treatment.Keywords: Lung cancer, Yes-associate protein, Transcriptional enhancer activation domain 4 (TEAD4), Peptide 17, Apoptosis

Highlights

Lung cancer, a potentially fatal disease, was first identified in 1761 [1]
The cell lysate was immunoprecipitated using anti-yesassociated protein (YAP) or antiTEAD4 antibody (Sigma, 1 : 1000), and the proteins were separated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDSPAGE) followed by immunoblotting
We found that the YAP/Transcriptional enhancer activation domain 4 (TEAD4) pathway was activated in both cancer cells

Summary

INTRODUCTION

A potentially fatal disease, was first identified in 1761 [1]. Until 1929, investigators believed that lung cancer initiation was closely associated with smoking [2]. Cells were cultured following the manufacturer’s instructions. RNA extraction and reverse transcription polymerase chain reaction (RT-PCR) analysis mirVana miRNA kit (Takara, China) was used to extract total RNA from the LLC and PC-9 cells following the manufacturer's instructions Of note, the internal control that we used was U6 small RNA. The LLC and PC-9 cells were lysed with RIPA buffer Primary antibodies, such as rat anti Ki67 (Santa Cruz, 1 : 500), rat anti Bax (Sigma, 1 : 1000), and mouse anti Bcl-2 (Santa Cruz, 1 : 1000), were integrated with the targeted protein by incubation at room temperature for 1-2 h. LLC and PC-9 cells were cultured in 6-well plates and supplemented with peptide 17 (5, 10, 20 and 40 nM). Differences with values of p < 0.05 were regarded as statistically significant

RESULTS

DISCUSSION

CONCLUSION

Conflict of Interest