Role of the stem cell-associated intermediate filament nestin in malignant proliferation of non-small cell lung cancer.
BACKGROUND
Nestin is associated with neoplastic transformation, but the mechanisms by which nestin contributes to invasion and malignancy of lung cancer remain unknown. Considering that proliferation is necessary for malignant behavior, we investigated the mechanism of nestin action in association with the proliferative properties of non-small cell lung cancer (NSCLC).


METHODS
Nestin expression was examined in NSCLC specimens and cell lines. Associations with clinicopathological features, including prognosis and proliferative markers, were evaluated. Effects of nestin knockdown on proliferation and the signaling pathways involved were further investigated.


RESULTS
Nestin was expressed in most cancer specimens and all the tumor cell lines analyzed. High nestin expression in malignant tissue was associated with high Ki-67 or PCNA levels and poor patient outcomes. Conversely, knockdown of nestin expression led to significant inhibition of tumor cell proliferation, decreased colony forming ability, and cell cycle G1 arrest. Furthermore, nestin knockdown resulted in inhibition of Akt and GSK3β activation.


CONCLUSIONS
Our data demonstrate that nestin expression in NSCLC cells is associated with poor prognosis of patients and tumor cell proliferation pathway. Downregulation of nestin efficiently inhibited lung cancer cell proliferation, which might be through affecting cell cycle arrest and Akt-GSK3β-Rb signaling pathway.
Department of Thoracic Surgery, the First Affiliated Hospital, Sun Yat-sen University, Guangzhou, Guangdong, China ; Department of Cardiothoracic Surgery, Huangpu Division of the First Affiliated Hospital, Sun Yat-sen University, Guangzhou, Guangdong, China ; Lung Cancer Research Center of Sun Yat-sen University, Guangzhou, Guangdong, China.
Zhenguang
Z
Chen
Center for Stem Cell Biology and Tissue Engineering, Sun Yat-sen University, Key Laboratory for Stem Cells and Tissue Engineering, Ministry of Education, Guangzhou, Guangdong, China.
Jiancheng
J
Wang
Department of Rehabilitation, the First Affiliated Hospital, Sun Yat-sen University, Guangzhou, Guangdong, China.
Lie
L
Cai
Department of Thoracic Surgery, the Fifth Affiliated Hospital, Sun Yat-sen University, Zhuhai, Guangdong, China.
Beilong
B
Zhong
Department of Thoracic Surgery, the First Affiliated Hospital, Sun Yat-sen University, Guangzhou, Guangdong, China ; Lung Cancer Research Center of Sun Yat-sen University, Guangzhou, Guangdong, China.
Honghe
H
Luo
Department of Medical Statistics and Epidemiology, School of Public Health, Sun Yat-sen University, Guangzhou, Guangdong, China.
Yuantao
Y
Hao
Center for Stem Cell Biology and Tissue Engineering, Sun Yat-sen University, Key Laboratory for Stem Cells and Tissue Engineering, Ministry of Education, Guangzhou, Guangdong, China.
Weihua
W
Yu
Guangdong Lung Cancer Institute, Guangdong General Hospital & Guangdong Academy of Medical Sciences, Guangzhou, Guangdong, China.
Binchao
B
Wang
Department of Thoracic Surgery, the First Affiliated Hospital, Sun Yat-sen University, Guangzhou, Guangdong, China ; Lung Cancer Research Center of Sun Yat-sen University, Guangzhou, Guangdong, China.
Chunhua
C
Su
Department of Thoracic Surgery, the First Affiliated Hospital, Sun Yat-sen University, Guangzhou, Guangdong, China ; Lung Cancer Research Center of Sun Yat-sen University, Guangzhou, Guangdong, China.
Yiyan
Y
Lei
Lung Cancer Research Center of Sun Yat-sen University, Guangzhou, Guangdong, China ; Department of Thoracic Surgery, Cancer Center, Sun Yat-sen University, Guangzhou, Guangdong, China.
Amos Ela
AE
Bella
Center for Stem Cell Biology and Tissue Engineering, Sun Yat-sen University, Key Laboratory for Stem Cells and Tissue Engineering, Ministry of Education, Guangzhou, Guangdong, China ; Department of Biochemistry, Zhongshan Medical School, Sun Yat-sen University, Guangzhou, Guangdong, China.
Andy Peng
AP
Xiang
Center for Stem Cell Biology and Tissue Engineering, Sun Yat-sen University, Key Laboratory for Stem Cells and Tissue Engineering, Ministry of Education, Guangzhou, Guangdong, China.
Tao
T
Wang
D002289:Carcinoma, Non-Small-Cell Lung / Q000235:genetics / Q000378:metabolism* / Q000473:pathology; D045744:Cell Line, Tumor; D049109:Cell Proliferation; D005260:Female; D059585:G1 Phase Cell Cycle Checkpoints / Q000235:genetics; D015972:Gene Expression Regulation, Neoplastic; D038362:Glycogen Synthase Kinase 3 / Q000378:metabolism; D006801:Humans; D015151:Immunoblotting; D007150:Immunohistochemistry; D019394:Ki-67 Antigen / Q000378:metabolism; D008175:Lung Neoplasms / Q000235:genetics / Q000378:metabolism* / Q000473:pathology; D008297:Male; D008856:Microscopy, Fluorescence; D008875:Middle Aged; D014411:Neoplastic Stem Cells / Q000378:metabolism* / Q000473:pathology; D064231:Nestin / Q000235:genetics / Q000378:metabolism*; D011379:Prognosis; D018809:Proliferating Cell Nuclear Antigen / Q000378:metabolism; D051057:Proto-Oncogene Proteins c-akt / Q000378:metabolism; D034622:RNA Interference; D020133:Reverse Transcriptase Polymerase Chain Reaction
D019394:Ki-67 Antigen; D064231:Nestin; D018809:Proliferating Cell Nuclear Antigen; D051057:Proto-Oncogene Proteins c-akt; D038362:Glycogen Synthase Kinase 3