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Down-regulated expression of family with sequence similarity 3, member B (FAM3B), in oral squamous cell carcinoma  Masashi Shiiba, Shunsaku Ishige, Yasuhiro.

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Presentation on theme: "Down-regulated expression of family with sequence similarity 3, member B (FAM3B), in oral squamous cell carcinoma  Masashi Shiiba, Shunsaku Ishige, Yasuhiro."— Presentation transcript:

1 Down-regulated expression of family with sequence similarity 3, member B (FAM3B), in oral squamous cell carcinoma  Masashi Shiiba, Shunsaku Ishige, Yasuhiro Saito, Toshihiro Shimizu, Yasuyuki Minakawa, Atsushi Kasamatsu, Katsunori Ogawara, Katsuhiro Uzawa, Hideki Tanzawa  Oral Science International  Volume 9, Issue 1, Pages 9-16 (May 2012) DOI: /S (12) Copyright © 2012 Japanese Stomatological Society Terms and Conditions

2 Fig. 1 Family with sequence similarity 3, member B (FAM3B) mRNA expression in oral squamous cell carcinoma (OSCC)-derived cell lines. FAM3B mRNA levels were analyzed in OSCC-derived cells and human normal oral keratinocytes (HNOKs) by real-time quantitative reverse transcriptase-polymerase chain reaction analysis. Significant down-regulation of FAM3B mRNA was observed in the five OSCC-derived cell lines, as compared with HNOKs (*p<0.05; Mann–Whitney U-test). Data are expressed as means±SEM of triplicate experiments. Oral Science International 2012 9, 9-16DOI: ( /S (12) ) Copyright © 2012 Japanese Stomatological Society Terms and Conditions

3 Fig. 2 Family with sequence similarity 3, member B (FAM3B) protein expression in oral squamous cell carcinoma (OSCC)-derived cell lines. FAM3B protein expression in OSCC-derived cell lines was analyzed with Western blot analysis. FAM3B and β-actin protein were detected by rabbit anti-FAM3B antibody and mouse anti-β-actin antibody, respectively (A). FAM3B protein signals were normalized against β-actin expression, and FAM3B expression levels of OSCC-derived cell lines are shown as relative values vs. human normal oral keratinocytes (HNOKs) (B). The results showed FAM3B protein expression is significantly diminished in OSCC-derived cell lines when compared with HNOKs. Oral Science International 2012 9, 9-16DOI: ( /S (12) ) Copyright © 2012 Japanese Stomatological Society Terms and Conditions

4 Fig. 3 Expression profile of family with sequence similarity 3, member B (FAM3B) mRNA in primary oral squamous cell carcinoma (OSCC) samples. Down-regulated expression of FAM3B mRNA was validated against primary OSCC samples using real-time quantitative reverse transcriptase-polymerase chain reaction analysis. Relative mRNA expression levels in normal oral tissues and primary OSCCs ranged from to (median, 0.262) and to (median, 0.028), respectively (p=0.0047; paired t-test). Oral Science International 2012 9, 9-16DOI: ( /S (12) ) Copyright © 2012 Japanese Stomatological Society Terms and Conditions

5 Fig. 4 Immunohistochemical (IHC) analysis of family with sequence similarity 3, member B (FAM3B) protein expression in primary oral squamous cell carcinoma (OSCC) samples. Representative results of IHC analysis for FAM3B in normal oral tissue (A, B) and primary OSCC (C, D). Original magnification: ×100 (A, C) and ×400 (B, D). Positive immunoreactions for FAM3B in normal oral tissue were detected in cytoplasm. Primary OSCC tissues exhibited faint FAM3B protein expression. FAM3B IHC scores are shown (E). FAM3B IHC scores for OSCCs and normal oral tissues range from 24.0 to (median, 60.0) and 93.0 to (median, 152.5), respectively. FAM3B protein expression levels in OSCCs were significantly lower than those in normal oral tissues (p<0.001; Mann–Whitney U-test). Oral Science International 2012 9, 9-16DOI: ( /S (12) ) Copyright © 2012 Japanese Stomatological Society Terms and Conditions

6 Fig. 5 Predicted target genes of microRNAs (miRNAs). Predicted target genes of miR-181a, miR-181b, miR-200b, and miR-200c are shown. Analysis was performed using publicly available algorithms (TargetScan and microRNA.org). The results suggested that FAM3B mRNA is the target gene of these 4 miRNAs. Oral Science International 2012 9, 9-16DOI: ( /S (12) ) Copyright © 2012 Japanese Stomatological Society Terms and Conditions

7 Fig. 6 Expression profiles of microRNAs (miRNAs) in oral squamous cell carcinoma (OSCC)-derived cell lines. MiRNA microarray was performed to identify crucial miRNAs in OSCC. The expression values for miR-181a, miR-181b, miR-200b, and miR-200c were greater in HSC-2, HSC-3, and Sa3 cells, as compared with human normal oral keratinocytes (HNOKs). Oral Science International 2012 9, 9-16DOI: ( /S (12) ) Copyright © 2012 Japanese Stomatological Society Terms and Conditions

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