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Expression of Derlin-1 and its effect on expression of autophagy marker genes under endoplasmic reticulum stress in lung cancer cells.

Xu L, Wang ZH, Xu D, Lin G, Li DR, Wan T, Guo SL - Cancer Cell Int. (2014)

Bottom Line: Knockdown of Derlin-1 did not affect Beclin 1 and LC3II expression but disrupted the degradation of p62 under ER stress, which resulted in the blockage of autophagy flux.Furthermore, Derlin-1 and p62 were observed to interact under ER stress.Derlin-1 may function in tumour progression partially by interacting with p62.

View Article: PubMed Central - HTML - PubMed

Affiliation: Department of Respiratory Medicine, The First Affiliated Hospital of Chongqing Medical University, Chongqing 400016, P.R. China.

ABSTRACT

Background: Recent findings indicated that Derlin-1 has an important function in tumour progression. In this study, we aimed to determine whether Derlin-1 has an oncogene function as a cross-talk molecule with autophagy.

Methods: Cancer cells were treated with tunicamycin (TM) for 8 and 24 h. The expression of Derlin-1 and autophagy-related genes was determined by western blot. Autophagy was analysed by fluorescence microscopy after staining the cancer cells with monodansylcadaverine. The interaction between Derlin-1 and other proteins was identified using co-immunoprecipitation assay.

Results: Our study demonstrated high Derlin-1 expression levels in most non-small lung cancer cell lines. Derlin-1 expression was enhanced under endoplasmic reticulum (ER) stress. Previous studies revealed that TM triggers the initiation of autophagy by activating Beclin 1, converting LC3I to LC3II and degrading p62. Knockdown of Derlin-1 did not affect Beclin 1 and LC3II expression but disrupted the degradation of p62 under ER stress, which resulted in the blockage of autophagy flux. Furthermore, Derlin-1 and p62 were observed to interact under ER stress.

Conclusion: This study is the first report about the interaction between Derlin-1 and p62. Derlin-1 may function in tumour progression partially by interacting with p62.

No MeSH data available.


Related in: MedlinePlus

Detection of Derlin-1 and autophagy marker gene expression after transfection of Derlin-1 siRNA with or without TM in A549 cells using western blot. A) Detection of autophagy marker gene expression after transfection of Derlin-1 siRNA with or without TM in A549 cells. B) Detection of Derlin-1 expression after TM exposure in A549 cells. The results show no significant alteration in Beclin 1 and LC3II expression after Derlin-1 siRNA transfection, but the accumulation of p62 indicates a disruption in p62 degradation and impairment of autophagy flux.
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Figure 3: Detection of Derlin-1 and autophagy marker gene expression after transfection of Derlin-1 siRNA with or without TM in A549 cells using western blot. A) Detection of autophagy marker gene expression after transfection of Derlin-1 siRNA with or without TM in A549 cells. B) Detection of Derlin-1 expression after TM exposure in A549 cells. The results show no significant alteration in Beclin 1 and LC3II expression after Derlin-1 siRNA transfection, but the accumulation of p62 indicates a disruption in p62 degradation and impairment of autophagy flux.

Mentions: To confirm the effect of Derlin-1 on autophagy gene expression, we transfected the siRNA target to Derlin-1 following TM treatment in A549 cells, which have high endogenous levels of Derlin-1. As shown in Figure 3, no significant alteration in Beclin 1 and LC3II expression was observed after Derlin-1 siRNA transfection. However, p62 accumulation was observed, indicating the disruption of p62 degradation and impairment of autophagy flux.


Expression of Derlin-1 and its effect on expression of autophagy marker genes under endoplasmic reticulum stress in lung cancer cells.

Xu L, Wang ZH, Xu D, Lin G, Li DR, Wan T, Guo SL - Cancer Cell Int. (2014)

Detection of Derlin-1 and autophagy marker gene expression after transfection of Derlin-1 siRNA with or without TM in A549 cells using western blot. A) Detection of autophagy marker gene expression after transfection of Derlin-1 siRNA with or without TM in A549 cells. B) Detection of Derlin-1 expression after TM exposure in A549 cells. The results show no significant alteration in Beclin 1 and LC3II expression after Derlin-1 siRNA transfection, but the accumulation of p62 indicates a disruption in p62 degradation and impairment of autophagy flux.
© Copyright Policy - open-access
Related In: Results  -  Collection

License 1 - License 2
Show All Figures
getmorefigures.php?uid=PMC4061450&req=5

Figure 3: Detection of Derlin-1 and autophagy marker gene expression after transfection of Derlin-1 siRNA with or without TM in A549 cells using western blot. A) Detection of autophagy marker gene expression after transfection of Derlin-1 siRNA with or without TM in A549 cells. B) Detection of Derlin-1 expression after TM exposure in A549 cells. The results show no significant alteration in Beclin 1 and LC3II expression after Derlin-1 siRNA transfection, but the accumulation of p62 indicates a disruption in p62 degradation and impairment of autophagy flux.
Mentions: To confirm the effect of Derlin-1 on autophagy gene expression, we transfected the siRNA target to Derlin-1 following TM treatment in A549 cells, which have high endogenous levels of Derlin-1. As shown in Figure 3, no significant alteration in Beclin 1 and LC3II expression was observed after Derlin-1 siRNA transfection. However, p62 accumulation was observed, indicating the disruption of p62 degradation and impairment of autophagy flux.

Bottom Line: Knockdown of Derlin-1 did not affect Beclin 1 and LC3II expression but disrupted the degradation of p62 under ER stress, which resulted in the blockage of autophagy flux.Furthermore, Derlin-1 and p62 were observed to interact under ER stress.Derlin-1 may function in tumour progression partially by interacting with p62.

View Article: PubMed Central - HTML - PubMed

Affiliation: Department of Respiratory Medicine, The First Affiliated Hospital of Chongqing Medical University, Chongqing 400016, P.R. China.

ABSTRACT

Background: Recent findings indicated that Derlin-1 has an important function in tumour progression. In this study, we aimed to determine whether Derlin-1 has an oncogene function as a cross-talk molecule with autophagy.

Methods: Cancer cells were treated with tunicamycin (TM) for 8 and 24 h. The expression of Derlin-1 and autophagy-related genes was determined by western blot. Autophagy was analysed by fluorescence microscopy after staining the cancer cells with monodansylcadaverine. The interaction between Derlin-1 and other proteins was identified using co-immunoprecipitation assay.

Results: Our study demonstrated high Derlin-1 expression levels in most non-small lung cancer cell lines. Derlin-1 expression was enhanced under endoplasmic reticulum (ER) stress. Previous studies revealed that TM triggers the initiation of autophagy by activating Beclin 1, converting LC3I to LC3II and degrading p62. Knockdown of Derlin-1 did not affect Beclin 1 and LC3II expression but disrupted the degradation of p62 under ER stress, which resulted in the blockage of autophagy flux. Furthermore, Derlin-1 and p62 were observed to interact under ER stress.

Conclusion: This study is the first report about the interaction between Derlin-1 and p62. Derlin-1 may function in tumour progression partially by interacting with p62.

No MeSH data available.


Related in: MedlinePlus