Osteopontin knockdown suppresses tumorigenicity of human metastatic breast carcinoma, MDA-MB-435.
Elevated expression of osteopontin (OPN), a secreted phosphoglycoprotein, is frequently associated with many transformed cell lines. Various studies suggest that OPN may contribute to tumor progression as well as metastasis in multiple tumor types. High levels of OPN have been reported in patients with metastatic cancers, including breast. We found that the expression of OPN corroborates with the aggressive phenotype of the breast cancer cells i.e. the expression of OPN is acquired as the breast cancer cells become more aggressive. To assess the role(s) of OPN in breast carcinoma, expression of endogenous OPN was knocked down in metastatic MDA-MB-435 human breast carcinoma cells using RNA interference. We targeted multiple regions of the OPN transcript for RNA interference, along with 'scrambled' and 'non-targeting siRNA pool' controls to distinguish between target-specific and potential off-target effects including interferon-response gene (PeIF2-alpha) induction. The OPN knockdown by shRNA suppressed tumor take in immunocompromised mice. The 'silenced' cells also showed significantly lower invasion and migration in modified Boyden chamber assays and reduced ability to grow in soft agar. Thus, in addition to the widely reported roles of OPN in late stages of tumor progression, these results provide functional evidence that OPN contributes to breast tumor growth as well.
Shevde
Lalita A
LA
University of South Alabama-Mitchell Cancer Institute, Mobile, AL 36688-0002, USA.
Samant
Rajeev S
RS
Paik
Jason C
JC
Metge
Brandon J
BJ
Chambers
Ann F
AF
Casey
Graham
G
Frost
Andra R
AR
Welch
Danny R
DR
D000818:Animals; D001943:Breast Neoplasms / Q000378:metabolism* / Q000473:pathology; D045744:Cell Line, Tumor; D018450:Disease Progression; D005260:Female; D006801:Humans; D051379:Mice; D008819:Mice, Nude; D009362:Neoplasm Metastasis / Q000235:genetics*; D053495:Osteopontin / Q000502:physiology*; D034741:RNA, Small Interfering / Q000494:pharmacology; D023041:Xenograft Model Antitumor Assays
D034741:RNA, Small Interfering; D053495:Osteopontin