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22 April 2020 : Clinical Research  

Prohibitin 2/PHB2 in Parkin-Mediated Mitophagy: A Potential Therapeutic Target for Non-Small Cell Lung Carcinoma

Han Zhang1ABCDEF, Chuntong Yin1ABCDEF, Xin Liu2ADEFG, Xue Bai2BCDE, Lei Wang2BCDE, Honglin Xu2BCDE, Jin Ju2BCDE, Linyou Zhang1ABCDEFG*

DOI: 10.12659/MSM.923227

Med Sci Monit 2020; 26:e923227

Abstract

BACKGROUND: Mitophagy, a selective autophagy process, plays various roles in tumors. Prohibitin 2 (PHB2) is an inner-mitochondrial membrane protein that participates in parkin-induced mitophagy. However, the role of PHB2 in non-small cell lung carcinoma (NSCLC) has not been previously reported.

MATERIAL AND METHODS: PHB2 protein or PHB2-mRNA in NSCLC and paired normal tissues was determined by Western blot, qRT-PCR, and immunohistochemical staining. Cell proliferation was detected by CCK-8 assay. Cell migration was evaluated by wound healing and transwell migration assays. A 3D live-cell confocal system was used to monitor autophagic flux. Mitochondrial autolysosomes were observed by transmission electron microscopy (TEM). Finally, we performed JC-1 assay to measure mitochondrial membrane potential (MMP).

RESULTS: The level of PHB2 was significantly increased in human NSCLC specimens compared to paired adjacent specimens. Inhibition of PHB2 expression attenuated mitophagy in A549 and H1299 cells, as indicated by decreased levels of LC3 II/I and parkin markers and increased level of p62 protein. Furthermore, the inhibition caused reduction in mitochondrial autolysosomes and autophagic flux, as shown by TEM and live-cell imaging, respectively. In addition, PHB2 inhibition caused a remarkable increase in MMP and suppressed the proliferation and migration of A549 and H1299 cells.

CONCLUSIONS: Our results suggest that downregulation of PHB2 reduced parkin-mediated mitophagy, which suppressed proliferation and migration of A549 and H1299 cells.

Keywords: Cell Migration Inhibition, Membrane Proteins, Mitochondrial Degradation, Aged, Aged, 80 and over, Apoptosis, Autophagy, Membrane Potential, Mitochondrial, Middle Aged, Mitochondria, Prohibitins, Repressor Proteins, Ubiquitin-Protein Ligases

Errate

Med Sci Monit 2022; 28:e937023     https://medscimonit.com/abstract/index/idArt/937023

The authors requested replacing Figure 2 as there was an error. The details of this error are as follows: Representative picture of transwell migration assay, A549 control group (Figure 2E). Representative picture of wound healing assay, 24h, A549 negative control group (Figure 2C). The above pictures were repeated within their own group (control group and negative control group). Representative picture of wound healing assay, 0h, A549 si-PHB2 group (Figure 2C). The authors used the wrong picture during data handling. Changes do not influence the results of the paper. In the original experiment, H1299 and A549 cells were divided into 4 groups (Control, si-PHB2, +PHB2, and negative control). Transwell migration assay and wound healing assay were performed 5 times. In addition, these results have been repeated by another research group (PHB2 promotes tumorigenesis via RACK1 in non-small cell lung cancer, Theranostics 2021, Vol. 11, Issue 7).
Reference: Han Zhang, Chuntong Yin, Xin Liu, Xue Bai, Lei Wang, Honglin Xu, Jin Ju, Linyou Zhang. Prohibitin 2/PHB2 in Parkin-Mediated Mitophagy: A Potential Therapeutic Target for Non-Small Cell Lung Carcinoma.  Med Sci Monit. 2020; 26: e923227. DOI: 10.12659/MSM.923227
 
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Med Sci Monit 2022; 28:e938532

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Medical Science Monitor eISSN: 1643-3750
Medical Science Monitor eISSN: 1643-3750