Endostar, a modified endostatin inhibits non small cell lung cancer cell in vitro invasion through osteopontin-related mechanism.
In this study, we studied the inhibition of non small cell lung cancer (NSCLC) cells invasion by a recombinant human Endostar, a modified endostatin and the possible osteopontin-related mechanism. The results showed that Endostar significantly inhibited highly metastatic NSCLC (NCI-H460) cells in vitro invasion. ELISA demonstrated that reduction of osteopontin level in the medium by Endostar may be responsible for the inhibition of invasion. RT-PCR assay and western blot analysis revealed that the reduction of osteopontin was due to under-regulation of osteopontin expression. Furthermore, Endostar also inhibited osteopontin-induced less metastatic NSCLC (A549) cells invasion, indicating that Endostar may have other different osteopontin-related mechanism. In an adhesion assay, we found that Endostar reduced NCI-H460 cells binding to osteopontin. Flow cytometric analysis suggested that the reduction of adhesion may be related to under-regulation of its receptors (CD44v6 and alpha(V)beta(3) integrin) expression. Additionally, we found, via gelatin zymographic analysis, that osteopontin-induced the expression and activation of pro-matrix metalloproteinase (MMP)-2 and pro MMP-9 secreted from A549 cells were blocked upon Endostar treatment, indicating that Endostar may block osteopontin-mediated signal transduction pathways through MMP families. The above results indicate that Endostar may have an intrinsic non-angiogenesis-related antitumor activity through osteopontin-related mechanism against NSCLC, including osteopontin change and osteopontin signal transduction blockade. Tumor cell invasion is important for tumor metastasis, our findings suggest that it is probably a good strategy to put Endostar into treatment of NSCLC metastasis.
Ni
Qinggui
Q
Department of Pharmacology, China Pharmaceutical University, Nanjing 210009, China.
Ji
Hui
H
Zhao
Zhihui
Z
Fan
Xin
X
Xu
Chengyun
C
D000818:Animals; D000970:Antineoplastic Agents / Q000494:pharmacology*; D002289:Carcinoma, Non-Small-Cell Lung / Q000235:genetics / Q000378:metabolism* / Q000473:pathology*; D045744:Cell Line, Tumor; D003470:Culture Media; D015536:Down-Regulation / Q000187:drug effects; D043169:Endostatins / Q000494:pharmacology*; D004789:Enzyme Activation / Q000187:drug effects; D015972:Gene Expression Regulation, Neoplastic / Q000187:drug effects; D006801:Humans; D008175:Lung Neoplasms / Q000235:genetics / Q000378:metabolism* / Q000473:pathology*; D045726:Metalloproteases / Q000235:genetics / Q000378:metabolism; D053495:Osteopontin / Q000235:genetics / Q000378:metabolism*; D011994:Recombinant Proteins
D000970:Antineoplastic Agents; D003470:Culture Media; D043169:Endostatins; D011994:Recombinant Proteins; D053495:Osteopontin; D045726:Metalloproteases; C522911:endostar protein