Clinicopathological significance of RCAN2 production in gastric carcinoma

概要

Clinicopathological significance of RCAN2 production in gastric carcinoma
㸦⫶⒴࡟࠾ࡅࡿ RCAN2 ⏘⏕ࡢ⮫ᗋ⑓⌮Ꮫⓗព⩏㸧

᭹㒊 ⤖㸦་ṑ⸆ಖ೺Ꮫ◊✲⛉ ་ṑ⸆Ꮫᑓᨷ㸧

඲ᩥせ⣙
࠙⫼ᬒࠚ
⫶⒴ࡣᅜෆእࢆၥࢃࡎṚஸ⋡ࡢ㧗࠸⒴⭘࡛࠶ࡾࠊ᪂つデ᩿࠶ࡿ࠸ࡣ἞⒪⸆ࡢ㛤Ⓨࡀ㔜せ࡞ㄢ㢟
࡛࠶ࡿࠋ௨๓௚◊✲ᐊ࡜ࡢඹྠ◊✲࡟࠾࠸࡚ࠊKRAS ኚ␗ᆺ኱⭠⒴࡛ࡣ RCAN2 ࡢⓎ⌧ᢚไࢆ
௓ࡋ Calcineurin–NFATc (cytosolic nuclear factor of activated T cells) ⤒㊰ࡢ⬺ᢚไ࡛⣽⬊ቑ
Ṫࡀஹ㐍ࡍࡿࡇ࡜ࠊVEGF ࢆ௓ࡋ࡚⾑⟶᪂⏕ࢆᢚไࡍࡿࡇ࡜ࢆ᫂ࡽ࠿࡟ࡋሗ࿌ࡋࡓࠋRCAN2
ࡣࣜࣥࣃ⌫ࡢάᛶ໬ࡸ⾑⟶᪂⏕࡟㛵୚ࡋ࡚࠸ࡿ calcineurin-NFATc signaling pathway ࡟࠾࠸
࡚ calcineurin regulator ࡜ࡋ࡚ാ࠸࡚࠸ࡿࠋࡋ࠿ࡋ࡞ࡀࡽࠊRCAN2 ࡢ⫶⒴࡟࠾ࡅࡿᙺ๭ࡣ඲
ࡃゎᯒࡉࢀ࡚࠸࡞࠸ࠋᮏ◊✲ࡣࠊKRAS ኚ␗ᆺ኱⭠⒴࡟࠾࠸࡚⭘⒆ᢚไⓗ࡞ാࡁࢆࡋ࡚࠸ࡿ
RCAN2 ⺮ⓑࡀྠࡌᾘ໬⟶⒴࡛࠶ࡿ⫶⒴࡟࠾࠸࡚ࡣ࡝ࡢࡼ࠺࡞ാࡁࢆࡋ࡚࠸ࡿࡢ࠿ࢆ᫂ࡽ࠿࡟
ࡍࡿࡇ࡜ࢆ┠ⓗ࡜ࡋ࡚ࠊ⫶⒴࡟࠾ࡅࡿ RCAN2 ࡢⓎ⌧ཬࡧ⏕≀Ꮫⓗᶵ⬟ゎᯒࢆ⾜ࡗࡓࠋ
࠙⤖ᯝࠚ
⫶⒴ࡢእ⛉ⓗษ㝖⤌⧊ 207 ౛࡟ᑐࡋච␿ᰁⰍ࡛ RCAN2 ⺮ⓑࡢⓎ⌧ࢆ᳨ウࡋࡓ࡜ࡇࢁࠊ110
౛ (53%)ࡀ㝧ᛶࢆ♧ࡋࡓࠋRCAN2 ⺮ⓑࡢⓎ⌧࡜⮫ᗋ⑓⌮ᏛⓗᅉᏊ࡜ࡢ㛵ಀࢆゎᯒࡋࡓ࡜ࡇࢁࠊ
stage ࡢ㐍⾜ (p <0.0001)ࠊ࣊ࣜࢥࣂࢡࢱ࣮ࣆࣟࣜឤᰁ (p=0.0023)࡜ࡢ㛫࡟᭷ព࡞┦㛵ࢆㄆࡵࡓࠋ
Kaplan-Meier ἲࢆ⏝࠸ࡓணᚋ࡜ࡢ㛵㐃᳨ウ࡛ࡣࠊRCAN2 㝧ᛶ౛ࡣ㝜ᛶ౛࡟ẚ࡭᭷ព࡟ணᚋ୙
Ⰻ࡛࠶ࡗࡓ (p <0.0001)ࠋ༢ኚ㔞 ࠊከኚ㔞ゎᯒࢆ⏝࠸ࡓ᳨ウ࡟࠾࠸࡚ࠊRCAN2 ࡣ⊂❧ࡋࡓணᚋ
୙ⰋᅉᏊ࡛࠶ࡿࡇ࡜ࡀ♧ࡉࢀࡓࠋ⒴⭘ࡢᾐ₶࣭ቑṪ࡬ࡢ㛵୚ࡀ▱ࡽࢀ࡚࠸ࡿ✀ࠎࡢศᏊ࡜ࡢ㛵ಀ
ࢆච␿ᰁⰍࢆ⏝࠸᳨࡚ウࢆ⾜ࡗࡓ࡜ࡇࢁࠊRCAN2 㝧ᛶ౛ࡣ EGFR (p <0.0001)ࠊβ-catenin (p

<0.0014)ࠊMMP-7 (p <0.0001)ࠊLamininγ2 (p <0.0001)ࠊVEGF (p <0.0001)㝧ᛶ౛࡜᭷ព࡞┦
㛵ࡀㄆࡵࡽࢀࡓࠋࡉࡽ࡟ࠊRCAN2 ࡜ࡇࢀࡽࡢศᏊࡢ⭘⒆ෆ࡟࠾ࡅࡿඹᒁᅾࡀච␿ᰁⰍ࡛☜ㄆ࡛
ࡁࡓࠋRCAN2 ࡢୗὶ࡛࠶ࡿ NFAT ࡢ୰࡛ࡶ⫶⒴࡜ࡢ㛵㐃ࡀሗ࿌ࡉࢀ࡚࠸ࡿ NFATc3 ࡶࠊච␿
ᰁⰍࢆ⏝࠸ࡓⓎ⌧᳨ウ࡟ࡼࡾ RCAN2 ࡜ࡢ㛫࡟┦㛵㛵ಀࡀㄆࡵࡽࢀࡓ (p <0.0001)ࠋ
RCAN2 ࡢ⫶⒴⣽⬊࡟࠾ࡅࡿ⏕≀Ꮫⓗᶵ⬟ࢆ᫂ࡽ࠿࡟ࡍࡿ┠ⓗ࡛ࠊRCAN2 ࡢ㧗Ⓨ⌧ࡀࡳࡽࢀ
ࡓ MKN1 ࢆ⏝࠸࡚ si-RNA ࡟ࡼࡿ RCAN2 ࡢࣀࢵࢡࢲ࢘ࣥࢆ⾜࠸ࠊMTT-assay ࡟࡚ቑṪ⬟ࢆࠊ
invasion assay ࡟࡚ᾐ₶⬟ࢆホ౯ࡋࡓࠋRCAN2 ࢆࣀࢵࢡࢲ࢘ࣥࡋࡓ⫶⒴⣽⬊ࡣ negative
control siRNA ࡛ฎ⌮ࡋࡓ⫶⒴⣽⬊࡜ẚ㍑ࡋ࡚ቑṪ⬟ ( p < 0.01)ࠊᾐ₶⬟ (p < 0.01) ࡀ᭷ព࡟
ᢚไࡉࢀࡓࠋࡋࡓࡀࡗ࡚ RCAN2 ࡣ⫶⒴ࡢቑṪ⬟ࠊᾐ₶⬟ࢆಁ㐍ࡍࡿࡶࡢ࡜ぢ࡞ࡉࢀࡓࠋRCAN2
ࡀ⫶⒴ࡢቑṪ࡟㛵୚ࡍࡿศᏊᶵᵓࢆ☜ㄆࡍࡿࡓࡵࠊEGFR ࡢୗὶ⤒㊰࡛࠶ࡿ ERK-MAPK ⤒㊰ࠊ
PI3K-AKT ⤒㊰࡬ࡢᙳ㡪ࢆ Western blot ἲ࡟᳨࡚ウࡋࡓࠋRCAN2 ࢆࣀࢵࢡࢲ࢘ࣥࡋࡓ⫶⒴⣽
⬊࡛ࡣ negative control siRNA ࡛ฎ⌮ࡋࡓ⫶⒴⣽⬊࡜ẚ㍑ࡋ ERK ཬࡧ AKT ࡢࣜࣥ㓟໬ࡀᢚไ
ࡉࢀࡓࠋ௨ୖ࠿ࡽ RCAN2 ࡣࡇࢀࡽࡢࢩࢢࢼࣝ⤒㊰ࢆ௓ࡋ࡚⒴ࡢቑṪ࡟㛵୚ࡋ࡚࠸ࡿ࡜⪃࠼ࡽ
ࢀࡓࠋࡲࡓࠊRCAN2 ࡢࣀࢵࢡࢲ࢘ࣥ࡟࡜ࡶ࡞ࡗ࡚ NFATc3 ࡢࣜࣥ㓟໬ࡀᢚไࡉࢀࡓࠋḟ࡟
RCAN2 ࡢⓎ⌧ࡀప࠸⫶⒴⣽⬊ᰴ MKN45 ࢆ⏝࠸࡚ RCAN2 ࡢᙉไⓎ⌧ࢆ⾜ࡗࡓࠋRCAN2 ࡢᙉ
ไⓎ⌧࡟ࡼࡾࠊempty ࡜ẚ㍑ࡋ࡚⫶⒴⣽⬊ᰴࡢቑṪ⬟ ( p < 0.01)ࠊᾐ₶⬟ ( p < 0.01)ࡀ᭷ព࡟
ୖ᪼ࡋࡓࠋ
࠙⪃ᐹࠚ
௨ୖࡢ⤖ᯝ࠿ࡽࠊRCAN2 ࡣ⫶⒴࡟࠾࠸࡚㧗Ⓨ⌧ࡋࠊRCAN2 ࡢ㧗Ⓨ⌧ࡣ⊂❧ࡋࡓணᚋ୙Ⰻᅉ
Ꮚ࡛࠶ࡿࡇ࡜ࡀ᫂ࡽ࠿࡜࡞ࡗࡓࠋࡲࡓ RCAN2 ࡢⓎ⌧ࡣ⭘⒆ࡢ㐍ᒎ࡟㛵୚ࡍࡿ EGFRࠊβ-cateninࠊ
MMP-7ࠊ Lamininγ2ࠊVEGF ࡢⓎ⌧࡜┦㛵ࡋࠊERK-MAPK ⤒㊰ࠊPI3K-AKT ⤒㊰ࢆάᛶ໬
ࡋ࡚ቑṪࠊᾐ₶ࢆಁ㐍ࡍࡿྍ⬟ᛶࡀ♧ࡉࢀࡓࠋRCAN2 ࡣ⫶⒴ࡢ᪂つデ᩿ཬࡧ἞⒪ᶆⓗศᏊ࡜ࡋ
࡚ᮇᚅࡉࢀࡿࠋ