Signal Activation of Hepatitis B Virus–Related Hepatocarcinogenesis by Up-regulation of SUV39h1

概要

ָ Ғ ࿨ ช સ ช གྷ ༁

Signal Activation of Hepatitis B Virus–Related
Hepatocarcinogenesis

by

Up-regulation

of

SUV39h1
㸦SUV39h1 Ⓨ⌧ஹ㐍࡟ࡼࡿ B ᆺ⫢⅖࢘࢖ࣝࢫ㛵㐃⫢
Ⓨ⒴ࡢಙྕάᛶ໬㸧
The Journal of Infectious Diseases, 222(12):
2061–2070, 2020.

➉ෆ ὈỤ
㸦་ṑ⸆ಖ೺Ꮫ◊✲⛉ ་ṑ⸆Ꮫᑓᨷ㸧

㸯㸧⫼ᬒ㸬┠ⓗ
B ᆺ⫢⅖࢘࢖ࣝࢫ(HBV)ឤᰁᝈ⪅࡟࠾࠸࡚ࡣ㸪ᢠ࢘࢖ࣝࢫ⒪ἲ࡟ࡼࡿ HBV ቑṪࡢไᚚࡣྍ⬟࡛
࠶ࡿࡀ㸪⫢Ⓨ⒴ࡀၥ㢟࡛࠶ࡿ㸬HBV 㛵㐃⺮ⓑࡢ 1 ࡘ࡛࠶ࡿ HBx ⺮ⓑࡣᝏᛶ໬ࡢㄏᑟࡸ࣑ࢺࢥ
ࣥࢻࣜ࢔ᶵ⬟୙඲࡞࡝ࢆ௓ࡋ㸪⫢Ⓨ⒴࡟㛵୚ࡋ࡚࠸ࡿ࡜⪃࠼ࡽࢀ࡚࠸ࡿ㸬୍᪉㸪SUV39h1 ࡀࣄ
ࢫࢺࣥࡢ࣓ࢳࣝ໬㓝⣲࡜ࡋ࡚Ⓨぢࡉࢀ࡚௨㝆㸪ᵝࠎ࡞ࣄࢫࢺ࣓ࣥࢳࣝ໬㓝⣲ࡀྠᐃࡉࢀࠊྛ㓝⣲
ࡀ≉ᐃࡢࣄࢫࢺࣥ࡟࠶ࡿࣜࢪࣥṧᇶࢆ࣓ࢳࣝ໬ࡋ㸪ࢡ࣐ࣟࢳࣥᵓ㐀ࡀኚ໬ࡉࡏ㸪㑇ఏᏊࡢ㌿෗ࢆ
ไᚚࡋ࡚࠸ࡿࡇ࡜ࡀ᫂ࡽ࠿࡜࡞ࡗ࡚࠸ࡿࡀ㸪HBx ࡜ SUV39h1 ࡢ┦஫స⏝࡟㛵ࡋ࡚ࡣ୙᫂࡞Ⅼ
ࡀከ࠸㸬ᮏ◊✲࡛ࡣ㸪ࣄࢺ⫢⣽⬊⛣᳜୙඲࣐࢘ࢫ㸦uPA-SCID ࣐࢘ࢫ㸧ࢆ⏝࠸࡚ HBV ᣢ⥆ឤᰁ
࡟క࠸㸪⫢⤌⧊ࡢ⥙⨶ⓗ࡞㑇ఏᏊⓎ⌧ゎᯒ࠿ࡽᢳฟࡉࢀࡓ SUV39h1 㑇ఏᏊ࡟╔┠ࡋ㸪HBV ឤ
ᰁ࡜ SUV39h1 ࡜ࡢ┦஫స⏝ࢆゎᯒࡋ㸪⫢Ⓨ⒴࣓࢝ࢽࢬ࣒࡬ࡢᙳ㡪ࡸ HBV ቑṪ࡬ࡢᙳ㡪࡟ࡘ࠸
᳨࡚ウࡋࡓ㸬

㸰㸧᪉ἲ࠾ࡼࡧ⤖ᯝ
ྛ࣐࢘ࢫ࡟ 106 copy ࡢ HBV(genotype C)ࢆྵࡴࣄࢺ⾑Ύࢆ࣐࢘ࢫᑿ㟼⬦⤒⏤࡛᥋✀ᚋ㸪⤒᫬ⓗ
࡟ HBV DNA ࢆ real time PCR ࡛ ᐃࡋࡓ࡜ࡇࢁឤᰁ᪩ᮇ(ឤᰁ 3 ᪥┠)ࢆ㝖ࡁࡍ࡭࡚ࡢ⩌࡛ᐃ
㔞ྍ⬟࡛࠶ࡗࡓ㸬
ࡇࢀࡲ࡛⾜ࡗࡓ cDNA ࣐࢖ࢡࣟ࢔ࣞ࢖ࡸḟୡ௦ࢩ࣮ࢡ࢚ࣥࢫ࡟ࡼࡿ᳨ウ࡛ࡣ㸪
HBV ࡢ▷ᮇឤᰁ࡛ࡣ⫢⣽⬊ෆ࡟࠾ࡅࡿ SUV39h1 ࡢⓎ⌧ୖ᪼ࡣㄆࡵ࡞ࡽࢀ࡞࠿ࡗࡓ㸬ᮏ◊✲࡛
ࡣࡉࡽ࡟㛗ᮇࡢほᐹࡢࡓࡵឤᰁ 238 ᪥ࡲ࡛ࡢ࣐࢘ࢫ⫢⣽⬊ෆࡢ SUV39h1 Ⓨ⌧ࢆ real time PCR
࡜ḟୡ௦ࢩ࣮ࢡ࢚ࣥࢫࢆ⏝࠸࡚ᐃ㔞ࡋࡓ㸬ࣄࢺ⫢⣽⬊࡟࠾ࡅࡿ SUV39h1 Ⓨ⌧ࡣ HBV ឤᰁ 56
᪥ᚋࡲ࡛ኚ໬ࡋ࡞࠿ࡗࡓࡀ㸪ឤᰁ 238 ᪥ᚋࡢ࣐࢘ࢫ࡛ࡣ HBV 㠀ឤᰁࡲࡓࡣ▷ᮇឤᰁ࣐࢘ࢫ࡜ẚ
㍑ࡋ࡚ SUV39h1 ࡢ᭷ព࡞Ⓨ⌧ୖ᪼ࢆㄆࡵࡓ㸬
ձ

⫢⒴ᝈ⪅࡛ࡣ㸪⒴⤌⧊ࡢ SUV39h1 Ⓨ⌧ࡀ㧗ࡃ㸪ࣄࢫࢺࣥ H3-K9 ࢺ࣓ࣜࢳࣝ໬ࡀஹ㐍ࡋࡓ
⑕౛࡛ࡣ↓෌Ⓨ⏕Ꮡ⋡ࡀ᭷ព࡟ప࠸࡜ሗ࿌ࡉࢀ࡚࠸ࡿ㸬ࡑࡇ࡛ 5 ౛ࡢ⫢⒴ᡭ⾡⑕౛ࡢ⫢⤌
⧊ࢆ⏝࠸࡚ SUV39h1 ࡢ㑇ఏᏊⓎ⌧ࢆᐃ㔞ࡋࡓ࡜ࡇࢁ㸪⒴㒊࡛ࡢⓎ⌧㔞ࡣ㠀⒴㒊࡜ẚ㍑ࡋ
࡚⣙ 2.1 ಸஹ㐍ࡋ࡚࠾ࡾ㸪SUV39h1 ࡢ⣽⬊ෆⓎ⌧ࡢஹ㐍ࡀ⒴໬࡜㛵㐃ࡋ࡚࠸ࡿࡇ࡜ࡀ♧၀
ࡉࢀࡓ㸬

ղ

SUV39h1 ࡢ⒴㒊࡟࠾ࡅࡿⓎ⌧ஹ㐍ࡀ☜ㄆࡉࢀࡓࡇ࡜࠿ࡽ㸪 HBV 㛵㐃⫢⒴࡟࠾ࡅࡿ
SUV39h1 ࡢᙳ㡪࡟ࡘ࠸᳨࡚ウࡋࡓ㸬HBV 㛵㐃⫢⒴࡛ࡣ HBV ࢤࣀ࣒ࡼࡾ⏘⏕ࡉࢀࡿ HBx
⺮ⓑࡀ⫢Ⓨ⒴࡟㛵୚ࡍࡿࡇ࡜ࡀሗ࿌ࡉࢀ࡚࠾ࡾ㸪HBx ࡜ SUV39h1 ࡢ┦஫స⏝ࢆゎᯒࡋࡓ㸬
HBx ࠾ࡼࡧ SUV39h1 Ⓨ⌧ࣉࣛࢫ࣑ࢻࢆస〇ࡋ㸪HepG2 ⣽⬊࡟ transfection ࡋࡓᚋ㸪⣽⬊
ෆ࡟࠾ࡅࡿ୧⺮ⓑࡢ interaction ࢆゎᯒࡋࡓ㸬ච␿ỿ㝆ࡢ⤖ᯝ㸪୧⺮ⓑࡀ interaction ࡍࡿ
ࡇ࡜ࡀ☜ㄆࡉࢀࡓ㸬᭦࡟㸪୧⺮ⓑ㛫ࡀ interaction ࡍࡿ㡿ᇦࡢྠᐃࡢࡓࡵ㸪HBx ࠾ࡼࡧ
SUV39h1 ࡢ඲㛗ࡶࡋࡃࡣ㒊ศ㛗ࢆ⤌ࡳ㎸ࢇࡔࣉࣛࢫ࣑ࢻࢆస〇ࡋ㸪ྠᵝࡢ᳨ウࢆ⾜ࡗࡓ࡜
ࡇࢁ㸪HBx ࡣ SUV39h1 ࡢ 89㹼233 ␒┠ࡢ㡿ᇦ㸦SET domain 㡿ᇦ㸧
㸪SUV39h1 ࡣ㸪HBx
ࡢ 52㹼102 ␒┠ࡢ㡿ᇦ㸦co-activator domain 㡿ᇦ㸧࡜ interaction ࡍࡿࡇ࡜ࡀ᫂ࡽ࠿࡜࡞
ࡗࡓ㸬

ճ

⥆࠸࡚㸪HBx ࡜ SUV39h1 ඹⓎ⌧࡟ࡼࡿⓎ⒴࣓࢝ࢽࢬ࣒࡬ࡢᙳ㡪࡟ࡘ࠸᳨࡚ウࡋࡓ㸬ࡇࢀ
ࡲ࡛࡟ HBx ࡟ࡼࡾⓎ⌧ࡀஹ㐍ࡋ⫢Ⓨ⒴࡟ᐤ୚ࡍࡿࡇ࡜ࡀሗ࿌ࡉࢀ࡚࠸ࡿ ATF6㸪AFP㸪
GADD45a㸪DUSP1 ࡢ 4 㑇ఏᏊ࡟╔┠ࡋゎᯒࢆ⾜ࡗࡓ㸬SUV39h1 ࠾ࡼࡧ HBV ࢆ HepG2
⣽⬊࡟ transfection ᚋ㸪⣽⬊ෆࡢ mRNA Ⓨ⌧ኚ໬ࢆ☜ㄆࡋࡓ࡜ࡇࢁ㸪HBV ࡜ SUV39h1
ࡢ୧⺮ⓑࡢᏑᅾୗ࡛㸪ࡇࢀࡽ 4 㑇ఏᏊࡢ᭷ព࡞Ⓨ⌧ஹ㐍ࢆㄆࡵࡓ㸬᭦࡟㸪HBV 㛵㐃⺮ⓑⓎ
⌧ࣉࣛࢫ࣑ࢻࢆ⏝࠸࡚ྠᵝࡢ᳨ウࢆ⾜ࡗࡓ࡜ࡇࢁ㸪4 㑇ఏᏊࡣ HBx Ⓨ⌧᫬ࡢࡳⓎ⌧ஹ㐍ࢆ
ㄆࡵ㸪ࡉࡽ࡟ SUV39h1 ࢆඹⓎ⌧ࡍࡿࡇ࡜࡟ࡼࡾࡑࡢⓎ⌧㔞ࡢቑᙉࢆㄆࡵࡓ㸬

մ

୍᪉㸪HBx ࡀ SUV39h1 ࡢ࣓ࢳࣝࢺࣛࣥࢫࣇ࢙࣮ࣛࢮάᛶࢆቑᙉࡍࡿྍ⬟ᛶࢆ⪃៖ࡋ㸪HBx
࠾ࡼࡧ SUV39h1 ࡢ recombinant protein ࢆస〇ᚋ㸪HBx ࡢ᭷↓࡟ࡼࡿ Histone H3 ࣓ࢳ
ࣝ໬ࡢ⛬ᗘࡢኚ໬ࢆ in vitro methyltransferase assay ࡟᳨࡚ウࡋࡓ㸬 SUV39h1 Ꮡᅾୗ࡛
Histone H3 ࡢ࣓ࢳࣝ໬ࡀほᐹࡉࢀ㸪 SUV39h1 ࡜ HBx ࡢඹᏑᅾୗ࡛ࡑࡢ࣓ࢳࣝ໬ࡀቑᙉ
ࡉࢀࡿࡇ࡜ࡀ☜ㄆࡉࢀࡓ㸬௨ୖ࠿ࡽ㸪HBx ࡢ SUV39h1 ࡟ࡼࡿ Histon H3 ࡢ࣓ࢳࣝ໬ࢆಁ
㐍ࡍࡿྍ⬟ᛶࡀ♧၀ࡉࢀࡓ㸬

㸱㸧⪃ᐹ
SUV39h1 ࡜ HBx ࡢ┦஫స⏝ࡀ஫࠸࡟άᛶࢆ㧗ࡵ㸪HBx ࢆ௓ࡋࡓ⫢Ⓨ⒴ࢆㄏᑟࡋ࡚࠸ࡿࡇ࡜ࡀ
♧၀ࡉࢀࡓ㸬SUV39h1 ࡢ HBx ᶵ⬟άᛶ໬ࡢ࣓࢝ࢽࢬ࣒ࢆゎᯒࡍࡿࡇ࡜࡟ࡼࡾ㸪HBV 㛵㐃Ⓨ⒴
࣓࢝ࢽࢬ࣒ࡢゎ᫂࡞ࡽࡧ࡟᪂ࡓ࡞⫢⒴἞⒪ᶆⓗ࡜࡞ࡿྍ⬟ᛶࡀ♧၀ࡉࢀࡓ㸬