Fused polypeptide with DEF induces apoptosis of lung adenocarcinoma cells.
To analyze the effects of a new unknown peptide DEF on the growth of tumor cells, a fused polypeptide TAT-DV1-DEF was designed and synthesized. The lung adenocarcinoma cell line GLC-82 treated with TAT- DV1-DEF was analyzed with a cell counting kit 8, and the location of polypeptides in cells was observed under laser confocal microscopy. The efficiency of polypeptide transfection and changes in nuclear morphology were analyzed by flow cytometry and fluorescence microscopy, respectively. Finally, the mechanism of tumor cell growth inhibition was evaluated by Western blotting. We found that TAT-DV1-DEF could significantly inhibit the growth of the lung adenocarcinoma cell line GLC-82, but not the normal human embryonic kidney cell line HEK-293. Polypeptides were found to be mostly localized in the cytoplasm and some mitochondria. The efficiency of polypeptide transfection in the two cell types was approximately 99%. Apoptotic nuclei were observed under fluorescence microscopy upon treatment with polypeptides and DAPI staining. Western blot analyses indicated that the polypeptide inhibition of tumor cell growth was apoptosis dependent. In the present study, we demonstrated that fused polypeptides could induce apoptosis of the lung adenocarcinoma cell line GLC-82, indicating that the new unknown peptide DEF has antitumor effects.
Key Laboratory for Clinical Laboratory Diagnostics of Education Ministry of China and School of Clinical Diagnostic and Laboratory Medicine, Chongqing Medical University, Chongqing, China E-mail : zhiguangtu106@163.com.
Ai-Ling
AL
Liang
Ting-Ting
TT
Zhang
Ning
N
Zhou
Di-Nan
DN
Huang
Xin-Guang
XG
Liu
Yong-Jun
YJ
Liu
Zhi-Guang
ZG
Tu
D000230:Adenocarcinoma / Q000378:metabolism / Q000473:pathology*; D017209:Apoptosis; D015153:Blotting, Western; D002467:Cell Nucleus / Q000378:metabolism; D049109:Cell Proliferation; D002478:Cells, Cultured; D003593:Cytoplasm / Q000378:metabolism; D005434:Flow Cytometry; D015696:Gene Products, tat / Q000235:genetics; D057809:HEK293 Cells; D006801:Humans; D008175:Lung Neoplasms / Q000378:metabolism / Q000473:pathology*; D008928:Mitochondria / Q000378:metabolism; D010446:Peptide Fragments / Q000494:pharmacology*; D011993:Recombinant Fusion Proteins / Q000494:pharmacology*
D015696:Gene Products, tat; D010446:Peptide Fragments; D011993:Recombinant Fusion Proteins