ホ౯ሗ࿌᭩SHE ⣽⬊ᙧ㉁㌿᥮ヨ㦂ἲ㸦SHE CTA㸧ᙧ㉁㌿᥮ヨ㦂㈨ᩱ⦅⧩ጤဨ఍

ᖹᡂ 29 ᖺ㸦2017 ᖺ㸧3 ᭶ 31 ᪥

1

7

ᙧ㉁㌿᥮ヨ㦂㈨ᩱ⦅⧩ጤဨ఍

ጤဨ㛗

ὸ㔝ဴ⚽

㸦ඖ᪥ᮾ㟁ᕤᰴᘧ఍♫㸪⌧኱㜰ಙឡዪᏛ㝔▷ᮇ኱Ꮫ㸧

ጤ ဨ

⟄஭೺ᶵ

㸦᪥ᮏṑ⛉኱Ꮫ㸧ᒣᙳᗣḟ㸦୍⯡㈈ᅋἲே㣗ရ⸆ရᏳ඲ࢭࣥࢱ࣮ ⛙㔝◊✲ᡤ㸧໭ᮏᖾᏊ㸦ఫ཭໬Ꮫᰴᘧ఍♫㸧➟ᯇಇኵ㸦ⰼ⋤ᰴᘧ఍♫㸧ᒣᮏ⨾ె㸦࢔ࢫࢸࣛࢫ〇⸆ᰴᘧ఍♫㸧

2

8

┠ ḟ

1.

せ᪨

2.

ヨ㦂᪉ἲࡢ⛉Ꮫⓗጇᙜᛶ࡜つไヨ㦂ἲ࡜ࡋ࡚ࡢጇᙜᛶ

3.

ヨ㦂᪉ἲࡢࣉࣟࢺࢥࣝࡢጇᙜᛶ

4.

ࣉࣞࣂࣜࢹ࣮ࢩࣙࣥ◊✲࡟⏝࠸ࡽࢀࡓ≀㉁ࡢศ㢮࡜ጇᙜᛶ5.

ヨ㦂ࡢṇ☜ᛶࢆホ౯ࡍࡿࡓࡵ࡟⏝࠸ࡽࢀࡓ໬Ꮫ≀㉁ࡢ in vivo ཧ↷ࢹ࣮ࢱ࡞ࡽࡧ࡟ཧ↷ࢹ࣮ࢱసᡂࡢ
⤒⦋

6.

ヨ㦂ࢹ࣮ࢱ࡜⤖ᯝࡢ฼⏝ᛶ

7.

ヨ㦂᪉ἲࡢṇ☜ᛶ

8.

ヨ㦂᪉ἲࡢಙ㢗ᛶ

9.

ヨ㦂᪉ἲࡢࢹ࣮ࢱࡢ㉁10.

ヨ㦂᪉ἲ࡟㛵ࡍࡿ௚ࡢ⛉Ꮫⓗ࡞ሗ࿌11.

3Rs ࡬ࡢ㛵୚㸦ື≀⚟♴㠃࠿ࡽࡢጇᙜᛶ㸧12.

ヨ㦂᪉ἲࡢ᭷⏝ᛶ࡜㝈⏺

13.

⤖ㄽ

14.

ཧ⪃ᩥ⊩

3

9

Abbreviations
3Rs:

Reduction, Replacement, and Refinement

CTA:

Cell Transformation Assay

CGM:

Complete Growth Medium

CIM:

Cell Isolation Medium

DB-ALM: DataBase service on Alternative Methods to Animal Experimentation
DRF:

Dose Range Finding assay

DRP:

Detailed Review Paper

DMEM:

Dulbecco’s modified Eagle’s medium

ESAC:

ECVAM Scientific Advisory Committee

EURL ECVAM: European Union Reference Laboratory for Alternatives to Animal Testing
FBS:

Foetal Bovine Serum

GD:

Guidance Document

IARC:

International Agency for Research on Cancer

MoA:

Mode of Action

MTF:

Morphological Transformation Frequency

NTP:

National Toxicology ProgramOECD:

Organization for Economic Co-operation and Development

SHE:

Syrian Hamster Embryo

UVBCs:

Substances of Unknown or Variable Composition, Complex Reaction Products or Biological Materials

WoE:

Weight of Evidence

WNT:

Working Group of National Coordinators of the Test Guidelines Program

4

10

1.

せ᪨
⣽⬊ࡢᙧ㉁㌿᥮ヨ㦂㸦Cell Transformation Assay : CTA㸧ࡣࠊin vitro ࡟࠾࠸࡚ࠊ໬Ꮫ≀㉁ࢆ᭚㟢ࡍࡿࡇ࡜࡛㉳ࡁࡿࠕ⣽⬊ࡢᙧែࠖࡸࠕ⾲⌧ᙧ㉁ࠖࡢᝏᛶ໬ࢆᣦᶆ࡜ࡋ࡚⭘⒆ㄏⓎᛶࡢ᭷↓ࢆㄪ࡭ࡿヨ㦂ἲࡢ୍ࡘ࡛࠶ࡿࠋ
JaCVAM SHE ⣽⬊ᙧ㉁㌿᥮ヨ㦂㈨ᩱ⦅⧩ጤဨ఍ࡣࠊOECD ࢞࢖ࢲࣥࢫ 1)࡜ࡋ࡚ 2015 ᖺ 5 ᭶࡟බ⾲ࡉࢀࡓࢩࣜ࢔࣭ࣥࣁ࣒ࢫࢱ࣮⫾௘㸦Syrian Hamster Embryo: SHE㸧⣽⬊ࢆ⏝࠸ࡿ CTA ࡀ໬Ꮫ≀㉁ࡢࡀࢇཎᛶࡢ
ண࡟᭷⏝࠿ྰ࠿࡟ࡘ࠸᳨࡚ウࡋࠊࡑࡢ⤖ᯝࢆࡲ࡜ࡵࡓࠋSHE ⣽⬊ࢆ⏝࠸ࡿ CTA ࡢ≉ᚩ࡜ࡋ࡚ࡣࠊ⏝࠸ࡿ⣽⬊ࡀึ௦ᇵ㣴⣽⬊࡛࠶ࡿࠊᇶᮏⓗ࡞௦ㅰ⬟ࢆ⥔ᣢࡋ࡚࠸ࡿࠊᇵ㣴ᮇ㛫ࡣ㸯㐌㛫⛬ᗘࡢ▷ᮇ࡛࠶ࡿࠊヨ㦂㈝⏝ࡣẚ㍑ⓗᏳ౯࡛࠶ࡿࠊࡋ࠿ࡋࠊᙧ㉁㌿᥮ࡋࡓࢥࣟࢽ࣮㸦⣽⬊㞟ⴠ㸧ࢆぢᴟࡵࡿ࡟ࡣ༑ศ࡞ᩍ⫱࡜カ
⦎ࡀᚲせ࡛࠶ࡿࡇ࡜࡞࡝ࡀᣲࡆࡽࢀࡿࠋ
SHE CTA ࡣࠊヨ㦂᪉ἲࡸࡀࢇཎᛶ࡜ࡢ┦㛵࡟㛵ࡍࡿㄽᩥࡀከࡃฟ∧ࡉࢀࠊࡑࡢ᭷⏝ᛶ࡟ࡘ࠸࡚ࡢ㆟ㄽࡶከࡃ࡞ࡉࢀ࡚࠸ࡿࠋEuropean Union Reference Laboratory for Alternatives to Animal Testing㸦EURL ECVAM㸧
࡟ࡼࡗ࡚ࠊࡑࡢヨ㦂᪉ἲ࡟㛵ࡍࡿࣉࣞࣂࣜࢹ࣮ࢩࣙࣥ◊✲ࡀᐇ᪋ࡉࢀࡓࠋࡋ࠿ࡋࠊ⿕㦂≀㉁ࡢᩘࡀᑡ࡞
ࡃࠊࡋ࠿ࡶヨ㦂㛤ጞ᫬࡟ࡣ㠀㑇ఏẘᛶࡀࢇཎ≀㉁࡜⪃࠼ࡽࢀ࡚࠸ࡓ≀㉁ࡀ୍✀㢮ᏑᅾࡋࡓࡶࡢࡢࠊOECD࡟ࡼࡿ SHE CTA ᑓ㛛ᐙ఍㆟࡟ࡼࡿ෌ホ౯⤖ᯝ 2)࡛ࡣࠊ㠀㑇ఏẘᛶࡀࢇཎ≀㉁ࡣྵࡲࢀ࡚࠸࡞࠸࡜ࡢ⤖ㄽࢆᚓࡓࠋ᭱⤊ⓗ࡟ࡣ⿕㦂≀㉁ࡢᩘࢆቑࡸࡋ࡚ࡢṇᘧ࡞ࣂࣜࢹ࣮ࢩࣙࣥ◊✲ࡣᐇ᪋ࡉࢀ࡞࠿ࡗࡓࡓࡵࠊヨ㦂᪉ἲࡀࣂࣜࢹ࣮ࢺࡉࢀࡓ࡜ࡣゝ࠸㞴࠸ࠋ ࡋ࠿ࡋ࡞ࡀࡽࠊࡀࢇཎ≀㉁࠾ࡼࡧ㠀ࡀࢇཎ≀㉁ࢆ⏝࠸ࡓ◊✲࡟ࡼࡿ࡜ࠊSHE CTA ࡢヨ㦂᪉ἲࡢṇ☜ᛶ࡟ࡘ࠸࡚ࡣࠊື≀ࢆ⏝࠸ࡓࡀࢇཎᛶヨ㦂࡜ࡢ୍⮴⋡ࠊឤᗘࠊ≉␗ᗘࠊഇ㝜ᛶ⋡࠾ࡼࡧഇ㝧ᛶ⋡ࡀࠊᴫࡡࠊ
ࡀࢇཎ≀㉁ࢆ᳨ฟࡍࡿ㑇ఏẘᛶヨ㦂ࢹ࣮ࢱ࡜ྠ➼ཪࡣࡑࢀ௨ୖࡢ⤖ᯝࡀᚓࡽࢀ࠾ࡾࠊࡑࡢ᭷⏝ᛶࡣ㧗࠸࡜⪃࠼ࡽࢀࡿࠋ
࡞࠾ࠊᮏ㉁ⓗ࡟ࡣࠊ⣽⬊ࡀᙧ㉁㌿᥮ࡍࡿ࣓࢝ࢽࢬ࣒ࡣᮍࡔᮍゎ᫂࡞ⅬࡀከࡃࠊCTA ࢆࡶࡗ࡚ື≀ࢆ⏝࠸ࡿࡀࢇཎᛶヨ㦂࡟⨨ࡁ᥮࠼ࡿࡇ࡜ࡣ⌧ẁ㝵࡛ࡣ୙㐺ษ࡛࠶ࡿ࡜⪃࠼ࡽࢀࡿࠋ
ࡇࡢࡼ࠺࡟ SHE CTA ࡀᣢࡘᅛ᭷ࡢၥ㢟ⅬࡣᏑᅾࡍࡿࡀࠊ ໬Ꮫ≀㉁࡟ࡼࡿⓎࡀࢇᶵᵓࡢ㔜せ࡞ࢫࢸࢵࣉ࡛࠶ࡿ⣽⬊ࡢᙧ㉁㌿᥮⬟ࢆホ౯࡛ࡁࡿ᪉ἲ࡛࠶ࡾࠊᐇ㝿ࠊࡀࢇཎ≀㉁ࡢ᭚㟢࡟ࡼࡾᙧ㉁㌿᥮ࢥࣟࢽ࣮
ࡢᩘࡀቑ࠼ࡿ࡜࠸࠺஦ᐇࢆ⪃៖ࡍࢀࡤࠊࡀࢇཎ≀㉁ࢆ᳨ฟࡋ࡚࠸ࡿ࡜⪃࠼ࡽࢀࡿࠋ≉࡟ International
Agency for Research on Cancer (IARC) ࢢ࣮ࣝࣉ 1 ࠾ࡼࡧ 2A ࡟ศ㢮ࡉࢀࡿ໬Ꮫ≀㉁࡟࠾࠸࡚ࠊSHE CTA ࡜ࣄࢺࡢࡀࢇཎᛶ࡜࡟┦㛵ࡀ࠶ࡾࠊᵓ㐀┦㛵࡞࡝࡜⤌ࡳྜࢃࡏࡿࡇ࡜࡟ࡼࡾࠊࡀࢇཎᛶࢆ␲ࢃࡏࡿ໬Ꮫ≀㉁࡞࡝ࡢࢫࢡ࣮ࣜࢽࣥࢢ⣔࡜ࡋ࡚ᛂ⏝ࡍࡿࡇ࡜࡟ࡼࡾࠊື≀ᩘࡢ኱ᖜ࡞๐ῶࡀྍ⬟࡜࡞ࡾࠊ3Rs ࡢ⢭⚄࡟ࡶ㐺࠺ヨ㦂᪉ἲ࡛࠶ࡿ࡜⪃࠼ࡽࢀࡿࠋࡇࡢࡼ࠺࡞ほⅬ࠿ࡽࠊSHE CTA ࡣࡉࡽ࡟ࢹ࣮ࢱࢆ✚ࡳ㔜ࡡࡿࡇ࡜࡟ࡼࡾࠊ໬Ꮫ≀㉁ࡢࡀࢇཎᛶண

ヨ㦂ࡢ᭷ᮃ࡞୍ࡘࡢヨ㦂ἲ࡟࡞ࡿ࡜⪃࠼ࡽࢀࡿࠋ

5

11

2.

ヨ㦂᪉ἲࡢ⛉Ꮫⓗጇᙜᛶ࡜つไヨ㦂ἲ࡜ࡋ࡚ࡢጇᙜᛶ

1䠅 ⛉Ꮫⓗጇᙜᛶ୍⯡ⓗ࡟ in vitro ࡟࠾ࡅࡿ CTA ࡜ࡣࠊ໬Ꮫ≀㉁ࡢ᭚㟢࡟ࡼࡾᇵ㣴⣽⬊ࡀ୙ྍ㏫ⓗ࡟ᙧែⓗኚ໬ࢆ㉳ࡇࡍ࠿࡝࠺࠿ࢆㄪ࡭ࡿヨ㦂࡛࠶ࡿࠋSHE CTA ࡣࠊࢩࣜ࢔࣭ࣥࣁ࣒ࢫࢱ࣮⫾௘࡟⏤᮶ࡍࡿึ௦⣽⬊ࢆ෾⤖࣭ゎ෾ࡋࡓ 2 ௦┠ࡢ⣽⬊㸦ṇᖖ஧ಸయ⣽⬊㸧ࢆ⏝࠸࡚ࠊ┠ⓗ࡜ࡍࡿ໬Ꮫ≀㉁ࢆ in vitro ࡛᭚㟢ࡋࡓᚋࠊᙧែᙧ㉁㌿᥮ࢥࣟࢽ࣮ࡢฟ⌧⋡࡟ࡼࡗ࡚ࠊ໬Ꮫ≀㉁ࡢᙧ㉁㌿᥮⬟ࡢ᭷↓ࢆホ౯ࡍࡿヨ㦂ἲ࡛࠶ࡿࠋSHE ⣽⬊ࡢᙧែᙧ㉁㌿᥮ࡣࠊከẁ㝵ⓗ⭘⒆ᙧᡂ㐣⛬ࡢ᪩ᮇ࡟㉳ࡁࡿ⌧㇟࡟┦ᙜࡋ࡚࠾ࡾࠊᙧ㉁㌿᥮⣽⬊ࡣࠊ⥅௦ᇵ㣴ᚋࠊྠ⣔⤫ࡢື≀ࡸࢾ࣮ࢻ࣐
࢘ࢫ࡟ᑐࡋ࡚⣙ 50㸣ࠥ100㸣ࡢ๭ྜ࡛㐀⭘⒆ᛶ 3,4)ࢆ♧ࡍࡇ࡜ࡀ▱ࡽࢀ࡚࠸ࡿࠋ
OECD Detailed Review Paper31 5) 㸦௨ୗ DRP31㸧࡟ࡣࠊ」ᩘࡢ pH ࡢᇵ㣴ᾮ࡛ヨ㦂ࡉࢀࡓᩘከࡃࡢ໬Ꮫ≀㉁ࡢ SHE CTA ⤖ᯝࡀࡲ࡜ࡵࡽࢀࠊ㑇ఏẘᛶࡀࢇཎ≀㉁࠾ࡼࡧ㠀㑇ఏẘᛶࡀࢇཎ≀㉁ࡢ᭚㟢࡟ࡼࡾ㸪
SHE ⣽⬊ࡣᙧែᙧ㉁㌿᥮ࡍࡿࡇ࡜ࡀ♧ࡉࢀ࡚࠸ࡿࠋEURL ECVAM ࡣࠊᶆ‽ࣉࣟࢺࢥࣝࠊᢏ⾡⛣㌿ᛶࠊ࠾ࡼࡧ᪋タෆ෌⌧ᛶ࡞ࡽࡧ࡟᪋タ㛫෌⌧ᛶࢆホ౯ࡋࡓ⤖ᯝࠊࡑࡢ᭷⏝ᛶࢆㄆࡵ࡚࠸ࡿࠋࡲࡓࠊEURL ECVAMࡢᶆ‽ⓗࣉࣟࢺࢥࣝ࡜ DRP31 ࡟♧ࡉࢀࡓࣉࣟࢺࢥࣝ࡜ࡢ㛫࡟ࡣᇶᮏⓗ࡞ᕪ␗ࡣ࡞ࡃࠊࡇࢀࡽࡢᚓࡽࢀࡓ
ࢹ࣮ࢱࡣᅇ㢳ⓗホ౯࡟࠾࠸࡚ࠊཷࡅධࢀྍ⬟࡛࠶ࡿ࡜ࡉࢀࡓࠋ
⣽⬊ࡢᙧ㉁㌿᥮ࡢṇ☜࡞ศᏊ⏕≀Ꮫⓗ࣓࢝ࢽࢬ࣒ࡣ༑ศ࡟ࡣゎ᫂ࡉࢀ࡚࠸࡞࠸ࡢࡀ⌧≧࡛࠶ࡿࠋSHE⣽⬊ࡀᙧ㉁㌿᥮ࡍࡿ࣓࢝ࢽࢬ࣒࡟ࡣ୙᫂࡞Ⅼࡀከ࠸ࡀࠊࡑࡢ࣓࢝ࢽࢬ࣒ࢆゎᯒࡍࡿㄽᩥࡣᩘከࡃ࠶ࡾࠊ
ᙧ㉁㌿᥮ࡣ༢୍ࡢ࣓࢝ࢽࢬ࣒࡛ㄏᘬࡉࢀ࡞࠸ࡇ࡜ࡶ᫂ࡽ࠿࡟࡞ࡗ࡚ࡁࡓࠋࡍ࡞ࢃࡕࠊ⣽⬊࿘ᮇไᚚࢩࢫ
ࢸ࣒ࠊࢤࣀ࣒Ᏻᐃᛶࠊ࠾ࡼࡧศ໬࣭ቑṪ㐣⛬࡛ࡢ㑇ఏᏊⓎ⌧ࡢኚ໬ࡸ␗ᖖࡀᙧ㉁㌿᥮ࡢ୺ࡓࡿせᅉ࡜ࡋ࡚⪃࠼ࡽࢀ࡚࠸ࡿࠋࡇࢀࡽࡢ㐣⛬࡟ᙳ㡪ࢆཬࡰࡍ㑇ఏⓗኚ␗ࡣ┤᥋ⓗ࡞㑇ఏẘᛶస⏝࡜⪃࠼ࡽࢀ࡚࠸ࡿࠋ
ࡉࡽ࡟ࠊDNA ࡢ㧗࣓ࢳࣝ໬࠶ࡿ࠸ࡣప࣓ࢳࣝ໬ࠊࣄࢫࢺࣥಟ㣭࠾ࡼࡧࢾࢡࣞ࢜ࢯ࣮࣒ࣜࣔࢹࣜࣥࢢ࡟ࡼ
ࡿ㑇ఏᏊⓎ⌧ࡢኚ໬ࡸ㑇ఏⓗ୙Ᏻᐃᛶࡀࠊ࢚ࣆࢪ࢙ࢿࢸࢵࢡ࡞ኚ໬ࢆㄏ㉳ࡋࠊᝏᛶᙧ㉁㌿᥮࡬࡜㐍⾜ࡉࡏࡿᘬࡁ㔠࡟࡞ࡿ࡜⪃࠼ࡽࢀ࡚࠸ࡿࠋ
ࡑࡢ௚ࡢ▱ぢ࡜ࡋ࡚ࠊ䞉୙Ṛ໬ࡸᝏᛶ໬ࡋࡓᙧ㉁㌿᥮⣽⬊࡟࠾ࡅࡿᚋᮇẁ㝵࡛ࡣࠊࡀࢇ㑇ఏᏊ ras ࡸ myc ࡟ᗈỗⓗࡶࡋࡃࡣ㒊఩≉␗ⓗ࡟ప࣓ࢳࣝ໬ࡀࡳࡽࢀ࡚࠸ࡿࡇ࡜ࠋ
䞉⣽⬊࿘ᮇࢳ࢙ࢵࢡ࣏࢖ࣥࢺ㑇ఏᏊࡢ࣓ࢳࣝ໬࡟ࡼࡿ㑇ఏᏊࡢⓎ⌧ᢚไࡸ p53 㑇ఏᏊࡢ✺↛ኚ␗࡜ྠᵝࠊ୙Ṛ໬ࢩࣜ࢔࣭ࣥࣁ࣒ࢫࢱ࣮⓶⭵⣽⬊࡟࠾࠸࡚ࠊࡀࢇᢚไ㑇ఏᏊ ink4a ࡸ ink4b ࡢᑐ❧㑇ఏᏊ࡟࣊ࢸࣟ
ࡲࡓࡣ࣍ࣔࡢḞኻࡀほᐹࡉࢀ࡚࠸ࡿࡇ࡜ࠋ
䞉ᙧ㉁㌿᥮ࡋࡓ SHE ⣽⬊ᰴ࡛ࡣ⣽⬊࿘ᮇࡢ G2 ࢳ࢙ࢵࢡ࣏࢖ࣥࢺ࡟㞀ᐖࡀ⏕ࡌ࡚࠸ࡿࡇ࡜ࠋ
䞉௚ࡢ⣽⬊ࢆᙧ㉁㌿᥮ࡉࡏࡿࡼ࠺࡞άᛶ໬ࣉࣟࢺ࢜ࣥࢥࢪ࣮ࣥ cph ࡀᝏᛶᙧ㉁㌿᥮⣽⬊࠿ࡽศ㞳ࡉࢀ࡚࠸ࡿࡇ࡜ࠋ࡞࡝ࡀሗ࿌ࡉࢀ࡚࠸ࡿࠋ
㠀㑇ఏẘᛶࡀࢇཎ≀㉁ࡶࡲࡓࠊከẁ㝵Ⓨࡀࢇ㐣⛬࡟㛵୚ࡋࠊṇᖖ⣽⬊ࢆᙧ㉁㌿᥮ࡉࡏࡿࡶࡢ࡜⪃࠼ࡽࢀ࡚࠸ࡿࠋࡍ࡞ࢃࡕࠊࢠࣕࢵࣉࢪࣕࣥࢡࢩࣙࣥ࡟ࡼࡿ⣽⬊㛫ࢥ࣑ࣗࢽࢣ࣮ࢩࣙࣥࢆ㜼ᐖࡍࡿ㓟໬ⓗࢫࢺ
612ࣞࢫࠊ⣽⬊ศ⿣ᅇᩘࡢቑຍࠊ࢔࣏ࢺ࣮ࢩࢫࡢῶᑡࠊࢳࣗࣈࣜࣥྜᡂ㜼ᐖࠊࢸ࣓࣮ࣟࣛࢮࡢάᛶ໬࡟ࡼࡿ⪁໬ࡢ㜼ᐖࠊࢩࢢࢼࣝఏ㐩㐣⛬ࡢ㜼ᐖࠊࡲࡓࠊ࣍ࣝࣔࣥ௓ᅾ㐣⛬ࡸ࣌ࣝ࢜࢟ࢩࢰ࣮࣒ࡢቑṪ࡟㛵୚ࡍࡿཷᐜ
య࡬ࡢ⤖ྜ࡞࡝࡛࠶ࡿࠋ
ࡇࡢࡼ࠺࡞⛉Ꮫⓗ▱ぢࡢከࡃࡀࠊࡀࢇ໬㐣⛬ุ࡛᫂ࡋ࡚࠸ࡿ஦ᐇ࡜ࡑࡢ୍㒊ࡀᩚྜࡍࡿࡇ࡜࡞࡝࠿ࡽࠊ
SHE CTA ࡢ⛉Ꮫⓗ࡞᳨ドࡣࠊ᏶඲࡛ࡣ࡞࠸ࡀࠊ⌧ᅾࡢศᏊ⏕≀ᏛⓗỈ‽࡟ᇶ࡙࠸࡚࠶ࡿ⛬ᗘ࡞ࡉࢀ࡚࠸
ࡿヨ㦂࡜⪃࠼ࡽࢀࡿࠋ

2䠅 つไୖࡢጇᙜᛶ
SHE CTA ࡣ༢୍ࡢ໬Ꮫ≀㉁࡟ࡘ࠸࡚ࡢࡳࣉࣞࣂࣜࢹ࣮ࢩࣙࣥ◊✲ࡀᐇ᪋ࡉࢀ࡚࠾ࡾࠊ࠸ࡃࡘ࠿ࡢ໬Ꮫ≀㉁࠿ࡽᵓᡂࡉࢀ࡚࠸ࡿ≀㉁ࠊUVCBsࠊᡈ࠸ࡣΰྜ≀࡟ࡘ࠸࡚ࡣࣂࣜࢹ࣮ࢩࣙࣥ◊✲ࡀᐇ᪋ࡉࢀ࡚࠸࡞
࠸ࡇ࡜ࢆ␃ពࡍ࡭ࡁ࡛࠶ࡿࠋࡇ࠺ࡋࡓ≧ἣࢆ⪃៖ࡋࡓୖ࡛ࠊOECD ࡢ࢞࢖ࢲࣥࢫࢻ࣓࢟ࣗࣥࢺ 1)ࡀ᫂☜࡟ࡋ࡚࠸ࡿࡇ࡜ࡣࠊࡀࢇཎᛶࢆホ౯ࡍࡿヨ㦂ᡓ␎ࡢ୍⎔࡜ࡋ࡚ࠊᡈ࠸ࡣࡀࢇཎᛶ࡟࠿࠿ࡿ WoE㸦ドᣐࡢ㔜ࡳ௜ࡅ㸧࡟ SHE CTA ⤖ᯝࢆ฼⏝ࡍࡿࡇ࡜ࡣ᭷⏝࡛࠶ࡿ࡜ࡋ࡚࠸ࡿࠋࡲࡓࠊSHE CTA ࡣ㑇ఏẘᛶࡀࢇཎ≀㉁࡟ᑐࡋ࡚㧗ឤཷᛶࢆࡶࡘࡀࠊ㠀㑇ఏẘᛶࡀࢇཎ≀㉁࡟ᑐࡋ࡚ࡣᮍࡔ㆟ㄽࡢవᆅࡀ࠶ࡾࠊࡼࡾከࡃࡢ᝟
ሗࡀồࡵࡽࢀ࡚࠸ࡿࡢࡶ஦ᐇ࡛࠶ࡿࠋᮏヨ㦂ἲࡀ OECD ヨ㦂ἲ࢞࢖ࢻࣛ࢖ࣥ࡜ࡋ࡚ᢎㄆࡉࢀ࡞࠿ࡗࡓ⌮⏤࡜ࡋ࡚ᣦ᦬ࡉࢀ࡚࠸ࡿࡇ࡜ࡣࠊᙧ㉁㌿᥮⣽⬊ࡢホ౯࡟㛵ࡋ࡚ᮏ㉁ⓗ࡞ၥ㢟ࡀࡑࡇ࡟ࡣᏑᅾࡍࡿࡇ࡜ࠊ
ࡍ࡞ࢃࡕࠊᙧ㉁㌿᥮࡟⮳ࡿศᏊ⏕≀Ꮫⓗస⏝ᶵᵓࡢ⌮ゎࡀ㝈ᐃⓗ࡛࠶ࡿࡇ࡜࡛࠶ࡿࠋࡲࡓ SHE CTA ࡢṇ☜ᛶศᯒ࡛⏝࠸ࡓ㠀㑇ఏẘᛶࡀࢇཎ≀㉁ࡀᑡ࡞࠿ࡗࡓࡇ࡜ 1,2)ࠊSHE CTA ࡢ⤖ᯝ࠿ࡽࡣࠊin vivo ࡟࠾ࡅࡿࡀࢇཎᛶࡢᙉࡉࠊ✀≉␗ᛶࡲࡓࡣ⤌⧊≉␗ᛶ࡞࡝ࡢ᝟ሗࡣᚓࡽࢀ࡞࠸ࡇ࡜࡞࡝ࡀᣦ᦬ࡉࢀ࡚࠸ࡿࠋ
ᮏヨ㦂ἲࡣ⌧⾜ࡢࡀࢇཎᛶヨ㦂࡟᏶඲࡟⨨ࡁ᥮ࢃࡿヨ㦂ἲ࡜ࡋ࡚ࡢ఩⨨࡙ࡅ࡛ࡣ࡞ࡃࠊ໬Ꮫ≀㉁➼ࡢࡀࢇཎᛶ࡟㛵ࡋ࡚ࡢ⾜ᨻⓗ࡞ุ᩿ࡀᚲせ࡟࡞ࡗࡓሙྜࠊ⿵ຓⓗ࡞ヨ㦂㸦౛࠼ࡤ Weight of Evidence㸸 WoE㸧࡜ࡋ࡚఩⨨࡙ࡅࡿࡇ࡜ࡀࡑࡢ฼⏝౯್ࢆ㧗ࡵࡿࡶࡢ࡜⪃࠼ࡽࢀࡿࠋ

3.

ヨ㦂᪉ἲࡢࣉࣟࢺࢥࣝࡢጇᙜᛶ

1㸧 ヨ㦂ἲࡢᴫ␎
SHE ⣽⬊ࡣ⫾⏕ 13 ᪥┠ࡢࢩࣜ࢔࣭ࣥࣁ࣒ࢫࢱ࣮⫾௘ࡢึ௦ᇵ㣴࠿ࡽᚓࡽࢀࡓṇᖖ஧ಸయ⣽⬊࡛ࠊ⸆≀௦ㅰ⬟ࡸ p53 άᛶࢆ᭷ࡍࡿࠋSHE ⣽⬊ࡣࠊ✀ࠎࡢࢱ࢖ࣉࡢ⣽⬊ᆺ࠿ࡽᵓᡂࡉࢀࡿࡓࡵࠊSHE CTA ࡣᰴ⣽⬊࠿ࡽ࡞ࡿ௚ࡢ CTA ἲ࡜ẚ㍑ࡋ࡚ࠊࡼࡾ⏕య࡟㏆࠸ṇᖖ⣽⬊࡟㉳ࡇࡿኚ໬ࢆᤊ࠼ࡿࡇ࡜ࡀ࡛ࡁࡿࠋከ✀ከᵝ࡞ࡀࢇཎ≀㉁ࢆ SHE ⣽⬊࡟᭚㟢ࡍࡿ࡜ࡲࡎᙧែᙧ㉁㌿᥮ࡀㄏᑟࡉࢀࠊᙧ㉁㌿᥮⣽⬊ࡣࠊ⥅௦ᇵ㣴ᚋࠊ኱㒊ศࡀ㐀⭘⒆ᛶ࣐࣮࣮࡛࢝࠶ࡿ㊊ሙ㠀౫ᏑᛶቑṪ⬟ࢆ⋓ᚓࡋࠊឤཷᛶࢆᣢࡘື≀࡟ᝏᛶ⭘⒆ࢆᙧᡂࡍ
ࡿࡇ࡜ࡀ▱ࡽࢀ࡚࠸ࡿࠋ
SHE CTA ࡣ⿕㦂≀㉁ࡢ᭚㟢࡟ࡼࡗ࡚ࠊከẁ㝵ⓗ࡟㐍⾜ࡍࡿ⭘⒆ᙧᡂ㐣⛬ࡢึᮇ࡟㉳ࡇࡿᙧ㉁㌿᥮ࢆ᳨ฟࡍࡿ in vitro ヨ㦂ἲ࡛࠶ࡿࠋᮏヨ㦂ἲ࡟ࡼࡗ࡚ࠊᚑ᮶ࡢ in vitro 㑇ఏẘᛶヨ㦂࠿ࡽࡣධᡭ࡛ࡁ࡞࠸⿕㦂≀㉁࡟₯ᅾࡍࡿ⣽⬊ᙧ㉁㌿᥮⬟ࢆ᳨ฟࡍࡿࡇ࡜ࡀ࡛ࡁࡿࠋ
SHE CTA ࡛ᚓࡽࢀࡓ⤖ᯝࢆໟᣓⓗ࡞ヨ㦂ᡓ␎ࡢ୍㒊ࡸ WoE ࡟ά⏝ࡍࢀࡤࠊ⿕㦂≀㉁ࡢࡶࡘ₯ᅾⓗᙧ
713

㉁㌿᥮⬟ࡢホ౯࡟౑࠼ࡿࡤ࠿ࡾ࡛࡞ࡃࠊSHE CTA ࡣࠊᚑ᮶ࡢື≀ࢆ⏝࠸ࡓࡀࢇཎᛶヨ㦂ࡢ୍㒊⨨ࡁ᥮࠼
㸦Replacement㸧࡞ࡽࡧ࡟ື≀ᩘࡢ๐ῶ㸦Reduction㸧࡟㈉⊩ࡍࡿࡇ࡜࡟࡞ࡾࠊࡑࡢព⩏ࡣ኱ࡁ࠸ࠋ

2㸧 ヨ㦂ἲࡢཎ⌮(1) SHE ⣽⬊ࡣ⫾⏕ 13 ᪥┠ࡢࢩࣜ࢔࣭ࣥࣁ࣒ࢫࢱ࣮⫾௘⣽⬊ࡢึ௦ᇵ㣴࠿ࡽᚓࡿࠋ㓝⣲࡛⫾௘⤌⧊ࢆᾘ໬ᚋࠊ⣽⬊ࢆ཰㞟ࡋᾮయ❅⣲୰࡛ಖᏑࡍࡿࠋ෾⤖⣽⬊ࡢ୍㒊ࡣࣇ࢕࣮ࢲ࣮⣽⬊㸦ᨭᣢ⣽⬊㸧࡟ࠊ௚ࡣᶆⓗ⣽⬊࡟౑⏝ࡍࡿࠋX ⥺↷ᑕ࡛ቑṪ⬟ࢆ୙ά໬ࡉࡏࡓ⣽⬊ࢆࣇ࢕࣮ࢲ࣮⣽⬊࡟⏝࠸ࡿࠋࣇ࢕
࣮ࢲ࣮⣽⬊ࡣᶆⓗ⣽⬊࡬ࡢᰤ㣴⿵⤥࡜⸆≀௦ㅰάᛶࢆࢧ࣏࣮ࢺࡍࡿ┠ⓗ࡛᧛✀ࡉࢀࠊᶆⓗ⣽⬊ࡣᙧ
ែᙧ㉁㌿᥮⬟ࢆㄪ࡭ࡿࡓࡵ࡟᧛✀ࡉࢀࡿࠋ(2) ᶆⓗ⣽⬊ࡣᙧᡂࡉࢀࡿࢥࣟࢽ࣮ࡢⓎ⫱ࢆ⪃៖ࡋ࡚ࠊCTA ࡢヨ㦂ᡂ❧ᇶ‽ࢆ‶ࡓࡍ࡟ᚲせ࡞ࢡ࣮ࣟࢽࣥࢢຠ⋡ࢆ㐩ᡂ࡛ࡁࡿࢡ࣮ࣟࣥᐦᗘ࡛ࣇ࢕࣮ࢲ࣮⣽⬊ᒙୖ࡬᧛✀ࡍࡿࠋ᧛✀ᚋࠊ⿕㦂≀㉁ࢆ 7 ᪥㛫᭚㟢ࡍࡿࠋࡑࡢᚋࠊ⣽⬊ࢆὙί࣭ᅛᐃࡋᰁⰍࡍࡿࠋࢹ࢕ࢵࢩࣗࡣࢥ࣮ࢻ໬ࡋࠊᐇయ㢧ᚤ㙾᳨ᰝ࡛ࢥࣟࢽ࣮ࡢᙧែⓗ⾲⌧ᆺࢆグ㘓ࡍࡿࠋ(3) ⣽⬊ẘᛶࡣࢥࣟࢽ࣮ᙧᡂຠ⋡࡜ࢥࣟࢽ࣮ࡢ኱ࡁࡉࡸ⣽⬊ᐦᗘࡢῶᑡ࠿ࡽホ౯ࡍࡿࠋグ㘓ࡋࡓࢥࣟࢽ
࣮ࡢ⥲ᩘ࡟ᑐࡍࡿᙧែᙧ㉁㌿᥮ࢥࣟࢽ࣮ᩘࡢ๭ྜࢆྛヨ㦂⃰ᗘ࡛⟬ฟࡍࡿࠋ⿕㦂≀㉁᭚㟢⩌ࡢ⥲ࢥࣟࢽ࣮ᩘ࡟ᑐࡍࡿᙧែᙧ㉁㌿᥮ࢥࣟࢽ࣮ᩘࡢ๭ྜࢆ⁐፹ᑐ↷⩌ࡢࡑࡢ๭ྜ࡜ẚ㍑ࡍࡿࠋ࡞࠾ࠊࢥࣟ
ࢽ࣮ࢧ࢖ࢬ࡜ᐦᗘ➼ࡢ≧ែࡣᚲせ࡟ᛂࡌ࡚グ㘓࡟ṧࡋࠊホ౯ࡢཧ⪃࡜ࡍࡿࡀࠊᙧ㉁㌿᥮⋡࡟ࡣᐤ୚ࡋ࡞࠸ࠋ

3㸧 ࣉࣟࢺࢥࣝࡢጇᙜᛶ
௨ୗࡢ㡯┠ࡢၥ㢟Ⅼࡀ᫂☜࡟ࡉࢀ࡚࠾ࡾࠊጇᙜᛶࡣᴫࡡ☜ಖࡉࢀ࡚࠸ࡿࠋ
(1)

ᐇ㦂⣔㸸
୍⭡ࡢࢩࣜ࢔࣭ࣥࣁ࣒ࢫࢱ࣮⫾௘࠿ࡽᚓࡽࢀࡓึ௦ᇵ㣴⣽⬊ࢆ౑⏝ࡍࡿࠋࡇࡢࡇ࡜ࡣ᭱ప࡛ࡶ 1 ༉ࡢዷፎࣁ࣒ࢫࢱ࣮ࡢᏳᴦṚࢆព࿡ࡍࡿࡀࠊ⣽⬊ࡢ㐺ษ࡞෾⤖ಖᏑ࡟ࡼࡗ࡚ࠊ1 ༉ࡢዷፎࣁ࣒ࢫࢱ࣮
࠿ࡽᑡ࡞ࡃ࡜ࡶ 50 ᅇศࡢ CTA ࢆᐇ᪋ࡍࡿࡢ࡟ᚲせ࡛༑ศ࡞⣽⬊ࢆᚓࡿࡇ࡜ࡀ࡛ࡁࡿࠋ

(2) SHE ⣽⬊࡜࢘ࢩ⫾௘⾑Ύ㸦FBS㸧ࡢࣟࢵࢺヨ㦂㸸
SHE ⣽⬊ࠊFBS ࡜ࡶ౑⏝࡟ඛ❧ࡗ࡚ࠊ᪂ࡋ࠸ࣟࢵࢺẖ࡟ CTA ࡟㐺ษ࠿࡝࠺࠿ࢆࢳ࢙ࢵࢡࡍࡿࠋ᭦
࡟ࠊ㐺ษ࡛࠶ࡿ࡜ุ᫂ࡋࡓ⣽⬊࡜ FBS ࢆ⤌ࡳྜࢃࡏ࡚ CTA ࢆᐇ᪋ࡋࠊᐃࡵࡽࢀࡓヨ㦂ᡂ❧ᇶ‽ࢆ‶ࡓࡍ࠿࡝࠺࠿ࡢヨ㦂ࢆ⾜࠺ࠋ(3) ⿕㦂≀㉁ࡢࣇ࢕࣮ࢲ࣮⣽⬊㞀ᐖᛶ☜ㄆ㸸(a) CTA ࡬ࡢ౑⏝࡟ඛ❧ࡗ࡚ࠊࣇ࢕࣮ࢲ࣮⣽⬊ࡔࡅࢆࢹ࢕ࢵࢩࣗ࡟᧛✀࣭ᇵ㣴ࡋࠊࢥࣟࢽ࣮ᙧᡂ⬟ࡢ࡞࠸ࡇ࡜ࢆ☜ㄆࡍࡿࠋ(b) ⿕㦂≀㉁࡛㑅ᢥⓗ࡟㞀ᐖࡉࢀࡓሙྜࢆ㝖࠸࡚ࠊ⿕㦂≀㉁᭚㟢⩌ࡢࢹ࢕ࢵࢩࣗୖࡢࣇ࢕࣮ࢲ࣮⣽⬊ࢆྍど࡛ࡁ࡞ࡅࢀࡤ࡞ࡽ࡞࠸ࠋ

8

14

(4) CTA ᢏ⾡ࡢ⇍㐩ᗘ☜ㄆヨ㦂㸸
CTA ᐇ᪋᪋タࡣᐃࡵࡽࢀࡓస⏝ᶵᗎࡢ␗࡞ࡿ㝧ᛶ࠾ࡼࡧ㝜ᛶᑐ↷≀㉁ࢆ⏝࠸࡚ CTA ࢆᐇ᪋ࡋࠊ
CTA ᢏ⾡ࡢ⇍㐩ᗘࢆ☜ಖࡍࡿࠋ(5) ౑⏝ࡍࡿᇵᆅࡢ pH㸦pH6.7 ࠾ࡼࡧ pH7.0)㸸
CTA ࡢᐇ᪋࡟ඛ❧ࡕࠊ㐺⏝ࡍࡿ pH ᮲௳࡛๓㏙ࡋࡓ CTA ᢏ⾡ࡢ⇍㐩ᗘ☜ㄆヨ㦂ࢆᐇ᪋ࡋࠊ⇍㐩ᗘࢆド᫂ࡍࡿࠋ
(6) ୧ pH 㛫ࡢ SHE CTA ࣉࣟࢺࢥࣝࡢᕪ␗㸸ᇵᆅ୰ࡢ FBS ⃰ᗘ࡟ᕪࡣ࠶ࡿࡀࠊࡑࢀ௨እ࡛ࡢᕪࡣㄆࡵࡽࢀ࡞࠸ࠋ
(7) ୧ pH 㛫࡟࠾ࡅࡿ SHE CTA ࣃࣇ࢛࣮࣐ࣥࢫࡢᕪࡢ᭷↓㸸࠸ࡎࢀࡢ pH ࡛ࡶ CTA ࡢ⤖ᯝࡣࠊឤᗘࠊ≉␗ᗘ࡜ࡶྠ⛬ᗘ࡛࠶ࡾࠊ኱ࡁ࡞ᕪࡣㄆࡵࡽࢀ࡞࠸ࠋ
(8) ヨ㦂ᡂ❧ᇶ‽㸸
CTA ࡢ⤖ᯝࡣࣉࣟࢺࢥ࡛ࣝつᐃࡋ࡚࠸ࡿヨ㦂ᡂ❧ᇶ‽࡟ᚑ࠸ࠊุᐃࡍࡿࠋ
(9) CTA ホ౯ࡢᐈほᛶࡢ☜ಖ㸸
CTA ࡟࠶ࡓࡗ࡚ࡣࠊ⿕㦂≀㉁ࡸᚓࡽࢀࡓࢹ࣮ࢱ࡟ࡘ࠸࡚ࢥ࣮ࢻ໬ࡋ࡚ホ౯ࡍࡿࠋ(10) ࢥࣟࢽ࣮ࡢ୺ほⓗࢫࢥ࢔ࣜࣥࢢ࡟ࡼࡿࢹ࣮ࢱࡢኚື㸸ࡇࡢᑐ⟇࡜ࡋ࡚ࠊ௨ୗࡢࡇ࡜ࢆᐇ᪋ࡍࡿࠋ
(a) ᪋タෆእࡢ CTA ⤒㦂⪅࡟ࡼࡿࢫࢥ࢔࣮ࣛࡢࢺ࣮ࣞࢽࣥࢢ(b) ✀ࠎࡢṇᖖ࠾ࡼࡧᙧ㉁㌿᥮ࢥࣟࢽ࣮ࡢ෗┿ぢᮏࡢ฼⏝㸦ࢫࢥ࢔ࣜࣥࢢ࡟࠶ࡓࡗ࡚ࡣࠊᖖ᫬෗┿ぢᮏࢆཧ⪃࡟ࡍࡿ㸧(c) ࢭ࢝ࣥࢻ࢜ࣆࢽ࢜ࣥ㸦௚ࡢࢫࢥ࢔࣮ࣛࡲࡓࡣࢫࢥ࢔⤒㦂⪅ࡢពぢࢆཧ⪃࡟ࡍࡿ㸧ࡸ௚ࡢࢫࢥ࢔
࣮ࣛ࡟ࡼࡿ஧㔜ࢫࢥ࢔ࣜࣥࢢ(d) ࢹ࢕ࢵࢩࣗࡢࢥ࣮ࢻ໬
(11) ⤖ᯝุᐃᇶ‽CTA ࡢ⤖ᯝࢆࣉࣟࢺࢥ࡛ࣝᐃࡵࡓ⤖ᯝุᐃᇶ‽࡟ᚑ࠸ࠊุᐃࡍࡿࠋ

4.

ࣉࣞࣂࣜࢹ࣮ࢩࣙࣥ◊✲࡟⏝࠸ࡽࢀࡓ≀㉁ࡢศ㢮࡜ጇᙜᛶࣉࣞࣂࣜࢹ࣮ࢩࣙࣥ◊✲ 6,7)࡟౪ࡉࢀࡓ 6 ≀㉁ࡢ࠺ࡕࠊ4 ≀㉁ࡀࡀࢇཎ≀㉁࡛ࠊࡑࡢ࠺ࡕࡢ 3 ≀㉁ࡣ㑇ఏẘᛶ≀㉁ࠊ1 ≀㉁ࡣ㠀㑇ఏẘᛶ≀㉁࡛࠶ࡿ࡜ヨ㦂㛤ጞ᫬࡟ࡣホ౯ࡉࢀ࡚࠸ࡓࡶࡢࡢࠊSHE CTA ᑓ㛛ᐙ఍㆟࡟ࡼࡿ෌᳨ウ⤖ᯝ࠿ࡽࠊ4 ≀㉁ࡣ඲࡚㑇ఏẘᛶ≀㉁࡛࠶ࡗࡓ࡜ุ᩿ࡉࢀࡓ 2)ࠋṧࡿ 2 ≀㉁ࡣ㠀㑇ఏẘᛶ㠀ࡀࢇཎ≀㉁࡛࠶ࡿࠋ඲య࡛ 6 ≀㉁࡜໬ྜ≀ᩘࡣᑡ࡞ࡃࠊ኱㒊ศࡀከ⎔ⰾ㤶᪘Ⅳ໬Ỉ⣲࡜ⰾ㤶᪘࢔࣑࡛ࣥࠊ⿕㦂≀㉁ࡢస⏝࣓࢝ࢽࢬ࣒࡟೫ࡾࡀࡳࡽࢀࡓࠋࡲࡓࠊ㝧ᛶ≀㉁ࡣ᫂ࡽ࠿࡞㝧ᛶ཯ᛂࢆ♧ࡍ໬ྜ≀ࡀከࡃ㸦4/6㸧ࠊࡀࢇཎᛶࡢ࠶࠸ࡲ࠸࡞཯ᛂࢆ♧ࡍ໬ྜ≀ࡣྵࡲࢀ࡞࠿ࡗࡓࡶࡢࡢࠊ㝜ᛶ≀㉁ࡶ 33㸣㸦2/6㸧
ࢆ༨ࡵ࡚࠾ࡾࠊ㑅ࡤࢀࡓ໬ྜ≀ࡣጇᙜ࡛࠶ࡿ࡜⪃࠼ࡽࢀࡓࠋ

915

㸺Genotoxic carcinogens㸼䞉Benzo(a)pyrene䞉2.4-Diaminotoluene䞉3-Methylcholanthrene䞉o-Toluidine㸨㸨ヨ㦂㛤ጞᙜ᫬ࡣ equivocal ࡲࡓࡣ inconclusive genotoxic carcinogen ࢆ non-genotoxic carcinogen ࡛࠶ࡿ࡜ゎ㔘ࡋ࡚࠸ࡓࠋ
㸺Non-genotoxic non-carcinogen㸼䞉Anthracene䞉Phthalic anhydride5.

ヨ㦂ࡢṇ☜ᛶࢆホ౯ࡍࡿࡓࡵ࡟⏝࠸ࡽࢀࡓ໬Ꮫ≀㉁ࡢ in vivo ཧ↷ࢹ࣮ࢱ࡞ࡽࡧ࡟
ཧ↷ࢹ࣮ࢱసᡂࡢ⤒⦋㠀㑇ఏẘᛶࡀࢇཎ≀㉁ࢆ᳨ฟࡍࡿ in vitro ヨ㦂ἲࡢ㛤Ⓨࡢ㔜せᛶࡀᣦ᦬ࡉࢀࡓࡇ࡜ࢆཷࡅࠊ1997 ᖺ࠿ࡽࠕNon-Genotoxic Carcinogens Detection: the Performance of In Vitro Cell Transformation Assaysࠖ࡜㢟ࡍࡿ
Detailed Review Paper ࡢసᡂࡀ㛤ጞࡉࢀࡓࠋࡑࢀࡣࠊSHE ⣽⬊ࠊBALB/c 3T3 ⣽⬊࠾ࡼࡧ C3H10T1/2 ⣽⬊ࢆ⏝࠸ࡿ CTA ࡟↔Ⅼࡀᙜ࡚ࡽࢀࡓࠋ2001 ᖺ࡟᭱ึࡢ᱌ࡀ The Working Group of National Coordinators of the
Test Guidelines Program㸦WNT㸧࡟ᅇぴࡉࢀࠊྛᅜࡢᑓ㛛ᐙ࠿ࡽࡢᗈ⠊ᅖ࡟ࢃࡓࡿࢥ࣓ࣥࢺࢆࡶ࡜࡟ᨵᐃ
∧ࡀసᡂࡉࢀࡓࡀࠊ᭱⤊໬࡟ࡣ⮳ࡽ࡞࠿ࡗࡓࠋ2002 ᖺ࡟ࡣࠊ➨ 2 ᱌ࡀᅇぴࡉࢀࠊ෌ࡧከᩘࡢࢥ࣓ࣥࢺࡀ
ฟࡉࢀࡓࡇ࡜࠿ࡽࠊࣇࣛࣥࢫ㸦SHE ⣽⬊㸧ࠊ᪥ᮏ㸦BALB/c3T3 ⣽⬊㸧ࠊ࢝ࢼࢲ㸦C3H10T1/2 ⣽⬊㸧ࡢ 3 ᅜࡀࠊࡑࢀࡒࢀࡢ⣽⬊ࢆ⏝࠸ࡓྛヨ㦂ἲ࡟ࡘ࠸࡚ᨵᐃసᴗࢆ⥅⥆ࡋ࡚㐍ࡵࡓ⤖ᯝࠊ2006 ᖺ࡟➨ 3 ᱌ࡀᥦ♧ࡉࢀࡓࠋ➨ 3 ᱌࡟ᑐࡍࡿࢥ࣓ࣥࢺ࡬ࡢ㆟ㄽ㸦࣡ࢩࣥࢺࣥ఍㆟㸧ࢆ⤒࡚➨ 4 ᱌ࡀసᡂࡉࢀࡓࠋࡇࡢ➨ 4 ᱌
ࠕDETAILED REVIEW PAPER ON CELL TRANSFORMATION ASSAYS FOR DETECTION OF CHEMICALCARCINOGENSࠖࡀ࠸ࢃࡺࡿ DRP31 ࡜ࡋ࡚ 2007 ᖺ 4 ᭶࡟㛤ദࡉࢀࡓ WNT19 ࡛ᢎㄆࡉࢀ᭱⤊໬ࡉࢀࡓࠋ
⾲㢟ࡀኚ᭦ࡉࢀ࡚࠸ࡿࡇ࡜࠿ࡽศ࠿ࡿࡼ࠺࡟ࠊࡇࡢ DRP31 ࡣࠊࡀࢇཎ≀㉁ࡢ᳨ฟ࡟࠾ࡅࡿ CTA ࡢព⩏ࢆ୺┠ⓗ࡜ࡋࠊ㠀㑇ఏẘᛶࡀࢇཎ≀㉁ࡢ in vitro ᳨ฟ⣔࡜ࡋ࡚ࡢศᯒࡣḟࡢㄢ㢟࡜ࡉࢀࡓࠋ
CTA ࡟㛵ࡋ࡚ࡣࠊୖ㏙ࡢ DRP31 సᡂ⤒⦋࡟࠶ࡿࡼ࠺࡟ SHE ⣽⬊௨እ࡟ BALB/c 3T3 ⣽⬊࠶ࡿ࠸ࡣ
C3H10T1/2 ࡞࡝ࡢ⣽⬊ࡀ⏝࠸ࡽࢀ࡚࠸ࡿࠋࡇࢀࡽࡢ⣽⬊࡛ࡢᙧ㉁㌿᥮⬟࡜ in vivo ࡀࢇཎᛶ࡜ࡢ┦㛵࡟ࡘ࠸࡚ࡣࠊDRP31 ࡀཧ↷ࡋᚓࡿࢹ࣮ࢱ㞟࡛࠶ࡿࠋ
DRP31 ࡛ࡣࠊࡆࡗṑ㢮࡟࠾ࡅࡿࡀࢇཎᛶヨ㦂ࢹ࣮ࢱࠊ㑇ఏẘᛶࢹ࣮ࢱ࡜ 3 ✀㢮ࡢ CTA ࢹ࣮ࢱࡀࡲ࡜ࡵ
ࡽࢀ࡚࠸ࡿࠋࡀࢇཎᛶヨ㦂ࢹ࣮ࢱ࡟㛵ࡋ࡚ࡣࠊIARC ࠾ࡼࡧ NTP ࡢࢹ࣮ࢱࡀ᥇⏝ࡉࢀ࡚࠸ࡿࠋ
DRP31 ࡛ࡣࠊSHE CTA ࡢ⤖ᯝ࡜ࡆࡗṑ㢮࡛ࡢࡀࢇཎᛶヨ㦂⤖ᯝࡢ୧᪉ࡀグ㍕ࡉࢀࡓ໬Ꮫ≀㉁ࡢᩘࡣ
245 ࡛࠶ࡿࠋࡑࡢෆヂࡣࠊࡀࢇཎ≀㉁ࡀ 175 ≀㉁ࠊ㠀ࡀࢇཎ≀㉁ࡀ 70 ≀㉁࡛࠶ࡿࠋෆヂࡣࠊ᭷ᶵ໬ྜ≀
ࡀ 190 ≀㉁㸦ࡀࢇཎ≀㉁㸸127 ≀㉁ࠊ㠀ࡀࢇཎ≀㉁㸸63 ≀㉁㸧ࠊ↓ᶵ໬ྜ≀ࡀ 55 ≀㉁㸦ࡀࢇཎ≀㉁㸸48≀㉁ࠊ㠀ࡀࢇཎ≀㉁㸸7 ≀㉁㸧࡛࠶ࡗࡓ㸦Appendix 1㸧ࠋ1016

SHE CTA ࢞࢖ࢻࣛ࢖ࣥࡢసᡂ㐣⛬࡟࠾࠸࡚ࠊࢹ࣮ࢱࡢ෌ホ౯ࡀᐇ᪋ࡉࢀࡓࠋChemical Carcinogenesis
Research Information System, NTP, IARC Monographs (1-106), PubMed,࠾ࡼࡧ CLP/GHS ࡢࢹ࣮ࢱ࣮࣋ࢫࢆ⏝࠸࡚ࠊSHE CTA ࡢࢹ࣮ࢱࡢࡳ࡞ࡽࡎࠊ㑇ఏẘᛶ࡟㛵ࡍࡿࢹ࣮ࢱ࠾ࡼࡧࡀࢇཎᛶࡢࢹ࣮ࢱ࡟ࡘ࠸࡚ࡶ෌ホ౯ࡉࢀࡓࠋࡑࡢ⤖ᯝࠊᑐ㇟࡜࡞ࡗࡓ 108 ≀㉁㸦ࡀࢇཎ≀㉁ 66 ≀㉁ࠊ㠀ࡀࢇཎ≀㉁ 42 ≀㉁㸧࡟ࡘ࠸࡚ࠊ෌ホ౯ࡉࢀࡓࢹ࣮ࢱࡀ♧ࡉࢀࡓ 2)ࠋ

6.

ヨ㦂ࢹ࣮ࢱ࡜⤖ᯝࡢ฼⏝ᛶࣉࣞࣂࣜࢹ࣮ࢩࣙࣥ◊✲࡟⏝࠸ࡓ 6 ≀㉁㸦㝧ᛶ 4 ≀㉁ࠊ㝜ᛶ 2 ≀㉁㸧ࡢࢹ࣮ࢱࡣࠊヨ㦂᪉ἲࡢಙ㢗ᛶࡢホ౯࡟⏝࠸ࡿࡇ࡜ࡀฟ᮶ࡿࠋ᪋タ㛫෌⌧ᛶࡢホ౯࡟ࡣ 6 ≀㉁඲࡚ࡢࢹ࣮ࢱࡀ⏝࠸ࡽࢀࡓࠋ୍᪉ࠊ᪋タෆ෌⌧ᛶࡢホ౯࡟ࡣ㝧ᛶᑐ↷≀㉁࡜ࡋ࡚౑ࢃࢀࡓ Benzo[a]pyrene ࡢࡳࡢ⤖ᯝࡀ⏝࠸ࡽࢀࠊ1 ᪋タ࡛ࡣࢥ࣮ࢻ໬ࡋ࡚⧞ࡾ㏉ࡋ 3 ᅇᐇ᪋㸦pH7.0 ἲ㸧ࡋࠊⰋዲ࡞⤖ᯝࡀᚓࡽࢀࡓࡀࠊ௚ࡢ 3 ᪋タ࡟ࡘ࠸࡚ࡣࠊẖᅇࢥ࣮ࢻ໬ࡏࡎ࡟㝧ᛶᑐ↷࡜ࡋ࡚ᐇ᪋ࡋࠊᚓࡽࢀࡓ್ࢆẚ㍑ࡋࡓ⤖ᯝ㸦pH6.7 ἲ㸧ࡋ࠿♧ࡉࢀ࡚࠸࡞࠸ࠋࡇࡢࡼ
࠺࡟ࠊᙉ࠸㝧ᛶ཯ᛂࢆ♧ࡍ 1 ໬ྜ≀ࡢࡳ࡛᪋タෆ෌⌧ᛶヨ㦂ࡀᐇ᪋ࡉࢀ࡚࠸ࡿࡇ࡜ࠊࡲࡓࠊྠ୍ࣉࣟࢺ
ࢥ࡛ࣝᐇ᪋ࡉࢀࡓヨ㦂࡛ࡣ࡞࠸ࡇ࡜࠿ࡽࠊ᪋タෆ෌⌧ᛶࢆホ౯ࡍࡿ࡟ࡣ༑ศ࡜ࡣゝ࠸㞴࠸ࠋ
DRP31 ࡛ホ౯ࡋࡓ≀㉁ࡢࢹ࣮ࢱࡣࠊヨ㦂᪉ἲࡢṇ☜ᛶࡢホ౯࡟⏝࠸ࡿࡇ࡜ࡀฟ᮶ࡿࠋ265 ≀㉁ࡢ࠺ࡕࠊ
157 ≀㉁ࡣࢻࣛࣇࢺࢸࢫࢺ࢞࢖ࢻࣛ࢖ࣥ㸦OECD, Feb. 2013㸧࡛ᐃࡵࡽࢀࡓ pH ࡸ⿕㦂≀㉁ฎ⌮᫬㛫࡛ᐇ᪋ࡉࢀ࡚࠸࡞࠸ࡓࡵࠊṇ☜ᛶࡢホ౯࠿ࡽ㝖እࡉࢀࡓࠋṧࡿ 108 ≀㉁ࡀࢻࣛࣇࢺࢸࢫࢺ࢞࢖ࢻࣛ࢖ࣥࡢࢡࣛ
࢖ࢸࣜ࢔࡟ᚑࡗ࡚෌ホ౯ࡉࢀࠊࡑࡢ⤖ᯝࡀヨ㦂᪉ἲࡢṇ☜ᛶࡢホ౯ 8)࡟౑ࢃࢀࡓࠋ

7.

ヨ㦂᪉ἲࡢṇ☜ᛶ
DRP31 ࡟ࡣࠊ㑇ఏẘᛶࢹ࣮ࢱࠊࡆࡗṑ㢮ື≀ࡢࡀࢇཎᛶヨ㦂ࢹ࣮ࢱ࡜ 3 ✀㢮ࡢ in vitro CTA ࢹ࣮ࢱࡀࡲ

࡜ࡵࡽࢀࠊࡀࢇཎᛶホ౯࡟࠾ࡅࡿ CTA ࢹ࣮ࢱࡢṇ☜ᛶ࡟ࡘ࠸࡚ሗ࿌ࡉࢀ࡚࠸ࡿࠋSHE CTA ࡢ⤖ᯝ࡟ࡘ࠸࡚ࡣࠊᇵ㣴ᾮࡢ pH ࡀ␗࡞ࡿヨ㦂ࢹ࣮ࢱࡀ⵳✚ࡉࢀ࡚࠸ࡿࡇ࡜࠿ࡽࠊpH6.7 ࠾ࡼࡧ pH7.0 ௨ୖࡢ 2 ࡘ࡟ศࡅ࡚ゎᯒࡉࢀ࡚࠸ࡿ㸦⾲ 1ࠊ⾲ 2㸧ࠋࡀࢇཎᛶホ౯࡟࠾ࡅࡿ SHE CTA ࠾ࡼࡧྛ✀㑇ఏẘᛶヨ㦂ࢹ࣮ࢱ࡟ࡘ࠸࡚ࡣࠊDRP31 ࡢ Annex ࡟ሗ࿌ࡉࢀ࡚࠾ࡾࠊࡑࢀࢆ⦅㞟ࡋࡓ⾲ 3 ࡟♧ࡋࡓ㏻ࡾࠊࡀࢇཎᛶ࡜ SHE CTA ࡢ୍⮴⋡ࡣྛ✀ࡢ㑇ఏẘᛶヨ㦂ࡢࡑࢀ࡜ྠ➼௨ୖ࡛࠶ࡾࠊഇ㝧ᛶ⋡࠾ࡼࡧഇ㝜ᛶ⋡ࡣప࠸࡜ゝ࠺⤖ᯝ࡜࡞ࡗ࡚࠸ࡿࠋࡲࡓࠊ⾲ 4 ࡟♧ࡍ㏻ࡾࠊྛ✀㑇ఏẘᛶヨ㦂࡜ CTA ࡟ඹ㏻ࡍࡿ≀㉁ࡢヨ㦂⤖ᯝ࡟ࡘ࠸࡚ࡀࢇཎᛶ࡜ࡢ୍⮴⋡ࢆࡳ࡚ࡶࠊSHE CTAࡣⰋዲ࡞⤖ᯝࢆ♧ࡋ࡚࠸ࡿࠋ

1117

⾲ 1 ࡀࢇཎᛶホ౯࡟࠾ࡅࡿ SHE CTA㸦pH6.7 ἲ㸧ࡢṇ☜ᛶ
In vivo ࡀࢇཎᛶࡀࢇཎ≀㉁㠀ࡀࢇཎ≀㉁SHE CTA

㝧ᛶ

36

5

㝜ᛶ

18

29

୍⮴⋡㸦Concordance㸧㸸(36+29)/(36+5+18+29)×100㸻65/88×100㸻74%ឤᗘ㸦Sensitivity㸧㸸

36/(36+18) ×100㸻36/54×100㸻67%≉␗ᗘ㸦Specificity㸧:

29/(5+29) ×100㸻29/34×100㸻85%ഇ㝜ᛶ⋡㸦False Negative㸧㸸

18/(36+18) ×100㸻18/54×100㸻33%ഇ㝧ᛶ⋡㸦False Positive㸧㸸

5/(5+29) ×100㸻5/34×100㸻15%

⾲ 2 ࡀࢇཎᛶホ౯࡟࠾ࡅࡿ SHE CTA㸦pH7.0 ἲ㸧ࡢṇ☜ᛶ
In vivo ࡀࢇཎᛶࡀࢇཎ≀㉁㠀ࡀࢇཎ≀㉁SHE CTA

㝧ᛶ

131

17

㝜ᛶ

11

33

୍⮴⋡㸦Concordance㸧㸸(131+33)/(131+17+11+33)×100㸻164/192×100㸻85%ឤᗘ㸦Sensitivity㸧㸸

131/(131+11) ×100㸻131/142×100㸻92%≉␗ᗘ㸦Specificity㸧:

33/(17+33) ×100㸻33/50×100㸻66%ഇ㝜ᛶ⋡㸦False Negative㸧㸸

11/(131+11) ×100㸻11/142×100㸻8%ഇ㝧ᛶ⋡㸦False Positive㸧㸸

17/(17+33) ×100㸻17/50×100㸻34%

⾲ 3 ࡀࢇཎᛶホ౯࡟࠾ࡅࡿ SHE CTA ࡜ྛ✀㑇ఏẘᛶヨ㦂ࡢẚ㍑
SHE CTA

Ames

ML

pH6.7 ἲpH7.0 ἲ୍⮴⋡㸦%㸧

74

85

52

72

ឤᗘ㸦%㸧

67

92

39.5

≉␗ᗘ㸦%㸧

8566

ഇ㝜ᛶ⋡㸦%㸧

33ഇ㝧ᛶ⋡㸦%㸧

HPRT

CA

MN

In vitro

In vivo

In vivo

78

62

60

58

86

80

65

57

58

79

26

70

59

68

55

8

60.5

14

20

35

43

42

15

34

21

74

30

41

32

45

ホ౯≀㉁⥲ᩘ

88

192

315

215

135

238

110

198

㠀ࡀࢇཎ≀㉁⋡㸦%㸧

39

26

29

30

22

30

29

25

Ames㸸᚟ᖐ✺↛ኚ␗ヨ㦂ࠊML㸸࣐࢘ࢫࣜࣥࣇ࢛࣮࣐ TK ヨ㦂ࠊ
HPRT㸸ᇵ㣴⣽⬊ࢆ⏝࠸ࡿ㑇ఏᏊ✺↛ኚ␗ヨ㦂ࠊCA㸸ᰁⰍయ␗ᖖヨ㦂ࠊMN㸸ᑠ᰾ヨ㦂1218

⾲ 4 ඹ㏻≀㉁ࡢࡀࢇཎᛶホ౯࡟࠾ࡅࡿ SHE CTA ࡜ྛ✀㑇ఏẘᛶヨ㦂ࡢẚ㍑
SHE

Ames

SHE

ML

SHE

HPRT

ホ౯≀㉁⥲ᩘ

242

166

111

㠀ࡀࢇཎ≀㉁⋡㸦%㸧

29

28

21

୍⮴⋡㸦%㸧

86

48

86

75

92

78

ឤᗘ㸦%㸧

90.5

36

88

86

96

79

≉␗ᗘ㸦%㸧

75

78

80

35

74

71

ഇ㝜ᛶ⋡㸦%㸧

10

64

12

14

4

21

ഇ㝧ᛶ⋡㸦%㸧

25

22

20

65

26

29

SHE

CA

SHE

CA

SHE

MN

In vitro

In vivo

In vivoホ౯≀㉁⥲ᩘ

180

78

154

㠀ࡀࢇཎ≀㉁ᩘ

29

29

26

୍⮴⋡㸦%㸧

86

62

86

66

83

56

ឤᗘ㸦%㸧

89

63

91

62.5

87

57

≉␗ᗘ㸦%㸧

77

59

75

71

71

54

ഇ㝜ᛶ⋡㸦%㸧

11

37

9

37.5

13

43

ഇ㝧ᛶ⋡㸦%㸧

23

41

25

29

29

46

SHE㸸SHE CTAࠊAmes㸸᚟ᖐ✺↛ኚ␗ヨ㦂ࠊML㸸࣐࢘ࢫࣜࣥࣇ࢛࣮࣐ TK ヨ㦂ࠊ
HPRT㸸ᇵ㣴⣽⬊ࢆ⏝࠸ࡿ㑇ఏᏊ✺↛ኚ␗ヨ㦂ࠊCA㸸ᰁⰍయ␗ᖖヨ㦂ࠊMN㸸ᑠ᰾ヨ㦂ࡇࡢࡼ࠺࡟ࠊOECD ࡣᩥ⊩ㄪᰝ࡟ࡼࡗ࡚ CTA ࡟࠾ࡅࡿࡀࢇཎ≀㉁ࡢண

ᛶࢆ᳨ドࡋࠊDRP31 ࡜ࡋ࡚ࡲ࡜ࡵࡓࠋࡑࡢ⤖ㄽ࡜ࡋ࡚ SHE CTA ࡢ࢞࢖ࢻࣛ࢖ࣥ໬ࡀ᥎ዡࡉࢀࡓࠋࡑࡢ⤖ᯝࠊุᐃἲࢆྵࡴヨ㦂ἲࡢᶆ‽໬࡞ࡽࡧ࡟࢞࢖ࢻࣛ࢖ࣥ໬࡟ྥࡅࠊOECD ࢞࢖ࢲࣥࢫ㸦OECD GD34ࠊ2005㸧9)࡟ᚑ࠸ࠊEURL ECVAM
୺ᑟ࡛ࣉࣞࣂࣜࢹ࣮ࢩࣙࣥ◊✲ࡀᐇ᪋ࡉࢀࡓࠋࡑࢀࡽࡢ⤖ᯝࢆホ౯ࡋࡓ EURL ECVAM Scientific Advisory
Committee㸦ESAC㸧ࡢሗ࿌᭩ 10)࡜ࡑࡢሗ࿌᭩ࢆཷࡅ࡚సᡂࡉࢀࡓ EURL ECVAM RECOMMENDATION11)ࡣ࡜ࡶ࡟ࠊSHE CTA ࡢ OECD ࢞࢖ࢻࣛ࢖ࣥ࡜ࡍࡿࡇ࡜ࢆ᥎ዡࡋࡓࠋࡇࡢࡼ࠺࡞⤒⦋ࢆ⤒࡚ࠊSHE CTA ࡢ࢞࢖ࢻࣛ࢖ࣥ᱌ࡀ 2013 ᖺ࡟බ⾲ࡉࢀࡓࠋࡋ࠿ࡋ࡞ࡀࡽࠊᥦ᱌ࡉࢀࡓヨ㦂ἲ࢞࢖ࢻࣛ࢖ࣥ᱌࡟ᑐࡋ࡚ࠊ㛵ಀྛᅜ࠿ࡽࡢࢥ࣓ࣥࢺ࡜ࡋ࡚ヨ㦂ἲ࢞࢖ࢻࣛ࢖ࣥ᱌ࡢุᐃᇶ‽࡜ DRP31 ࡢุᐃ⤖ᯝ࡜ࡢᩚྜᛶࡢ෌☜ㄆࡀồࡵࡽࢀࡓࠋ
ࡇࢀࢆཷࡅ࡚ࠊpH6.7 ἲ࡜ pH7.0 ἲ㸦pH7.0 ࢆ㉸࠼ࡿ᮲௳࡛ᐇ᪋ࡉࢀࡓࢹ࣮ࢱࡣ㝖እ㸧࡛ᐇ᪋ࡉࢀ࡚࠸
ࡿࢹ࣮ࢱࡢ෌ホ౯ࡀ⾜ࢃࢀࡓ 8)ࠋࡍ࡞ࢃࡕࠊpH6.7 ἲ࡛ࡣ 93 ≀㉁ࠊpH7.0 ἲ࡛ࡣ 42 ≀㉁࡟ࡘ࠸࡚෌ホ౯ࡀ⾜ࢃࢀࡓ㸦⾲ 5ࠊ⾲ 6㸧ࠋ

1319

⾲ 5 ࡀࢇཎᛶホ౯࡟࠾ࡅࡿ SHE CTA㸦pH6.7 ἲ㸧ࡢṇ☜ᛶ㸦෌ホ౯⤖ᯝ㸧In vivo ࡀࢇཎᛶSHE CTAࡀࢇཎ≀㉁㠀ࡀࢇཎ≀㉁

㝧ᛶ

42㸦6㸧

8㸦1㸧

㝜ᛶ

15㸦1㸧

26㸦0㸧

୙☜ᐃ㸦Equivocal㸧

1㸦0㸧

1㸦0㸧

୍⮴⋡㸦Concordance㸧㸸(42+26)/(42+8+15+26+1+1)×100㸻68/93×100㸻73%ឤᗘ㸦Sensitivity㸧㸸

42/(42+15+1) ×100㸻42/58×100㸻72%≉␗ᗘ㸦Specificity㸧:

26/(8+26+1) ×100㸻26/35×100㸻74%ഇ㝜ᛶ⋡㸦False Negative㸧㸸

15/(42+15+1) ×100㸻15/58×100㸻26%ഇ㝧ᛶ⋡㸦False Positive㸧㸸

8/(8+26+1) ×100㸻8/35×100㸻23%ᣓᘼෆࡢᩘ್ࡣ↓ᶵ≀㉁ࡢホ౯ᩘ⾲ 6 ࡀࢇཎᛶホ౯࡟࠾ࡅࡿ SHE CTA㸦pH7.0 ἲ㸧ࡢṇ☜ᛶ㸦෌ホ౯㸧
In vivo ࡀࢇཎᛶSHE CTAࡀࢇཎ≀㉁㠀ࡀࢇཎ≀㉁㝧ᛶ24㸦11㸧

1㸦0㸧

㝜ᛶ

6㸦2㸧

11㸦0㸧

୍⮴⋡㸦Concordance㸧㸸(24+11)/(24+1+6+11)×100㸻35/42×100㸻83%ឤᗘ㸦Sensitivity㸧㸸

24/(24+6) ×100㸻24/30×100㸻80%≉␗ᗘ㸦Specificity㸧:

11/(11+1) ×100㸻11/12×100㸻92%ഇ㝜ᛶ⋡㸦False Negative㸧㸸

6/(24+6) ×100㸻6/30×100㸻20%ഇ㝧ᛶ⋡㸦False Positive㸧㸸

1/(11+1) ×100㸻1/12×100㸻8%ᣓᘼෆࡢᩘ್ࡣ↓ᶵ≀㉁ࡢホ౯ᩘ
ࡲࡓࠊ2 ࡘࡢ᪉ἲ࡟ඹ㏻ࡢ≀㉁㸦ࡀࢇཎ≀㉁㸸14 ≀㉁ࠊ㠀ࡀࢇཎ≀㉁㸸4 ≀㉁㸧࡟ࡘ࠸࡚ࡢ⤖ᯝࡀẚ㍑
ࡉࢀࠊ14 ࡢࡀࢇཎ≀㉁ࡢෆ 1 ≀㉁㸦diethylstilbestrol㸧ࡢࡳ 2 ࡘࡢヨ㦂㛫࡛⤖ᯝࡀ୍⮴ࡋ࡚࠸࡞࠿ࡗࡓࡀࠊ
13 ≀㉁ࡣࡍ࡭࡚⤖ᯝࡀ୍⮴㸦12 ≀㉁㸸㝧ᛶࠊtitanium dioxide ࡢ 1 ≀㉁㸸㝜ᛶ㸧ࡋ࡚࠸ࡓࠋࡲࡓࠊ㠀ࡀࢇཎ≀㉁ࡢෆ 2 ≀㉁ࡣ 2 ヨ㦂࡛࡜ࡶ࡟㝜ᛶࠊṧࡾࡢ 2 ≀㉁ࡢ࠺ࡕࠊCaprolactam ࡣ pH6.7 ἲ࡛ࡣ㝜ᛶ࡛࠶ࡗ
ࡓࡀࠊpH7.0 ἲ࡛ࡣ㝧ᛶ࡛࠶ࡗࡓࠋࡲࡓࠊPhthalic anhydride ࡣ pH6.7 ἲ࡛ࡣヨ㦂᪋タ㛫࡛⤖ᯝ࡟ᕪࡀ࠶ࡾࠊ
pH7.0 ἲ࡛ࡣ㝜ᛶࡢ⤖ᯝ࡛࠶ࡗࡓࠋࡇࡢࡼ࠺࡟ࠊ2 ࡘࡢ᪉ἲ࡟ࡣ pH ࡢ㐪࠸ࡣ࠶ࡿࡀࠊ࡯ࡰྠ➼ࡢ⤖ᯝࡀᚓࡽࢀ࡚࠸ࡿ࡜⪃࠼ࡽࢀࡿࠋ
ࡀࢇཎ≀㉁ࢆ᭷ᶵ໬ྜ≀࡜↓ᶵ໬ྜ≀࡟ศࡅ࡚ࡑࡢ᳨ฟ⋡࡟ࡘ࠸࡚ࡶ᳨ウࡋ࡚࠸ࡿࠋpH6.7 ἲ࡟ࡘ࠸࡚
ࡣ↓ᶵ໬ྜ≀ࡀ 1 ≀㉁࡛࠶ࡾࠊ᭷⏝࡞᝟ሗ࡜ࡣ⪃࠼ࡽࢀ࡞࠸ࠋ୍᪉ࠊpH7.0 ἲ࡟ࡘ࠸࡚ࡣ᭷ᶵ໬ྜ≀ࡀ 17≀㉁ࠊ↓ᶵ໬ྜ≀ࡀ 13 ≀㉁࠶ࡾࠊ᳨ฟ⋡ࡣࡑࢀࡒࢀ 76.5% ࠾ࡼࡧ 84.6% ࡛࠶ࡗࡓࠋ᭦࡟ࠊSHE CTA ࡟࠾ࡅࡿ㠀㑇ఏẘᛶࡀࢇཎ≀㉁ࡢ᳨ฟ⋡࡟ࡘ࠸࡚ࡶホ౯ࡋ࡚࠸ࡿ 12)㸦⾲ 7㸧ࠋ

14

20

⾲ 7

SHE CTA ࡟࠾ࡅࡿ㠀㑇ఏẘᛶࡀࢇཎ≀㉁ࡢ᳨ฟ⋡㠀㑇ఏẘᛶࡀࢇཎ≀㉁᳨ฟ⋡㸦%㸧

SHE CTA

㝧ᛶ

14㸦1㸧

58

(pH6.7 ἲ)

㝜ᛶ

10㸦0㸧

42

ィ

24㸦1㸧

SHE CTA

㝧ᛶ

8㸦4㸧

67

(pH7.0 ἲ)

㝜ᛶ

4㸦1㸧

33

ィ

12㸦5㸧

ᣓᘼෆࡢᩘ್ࡣ↓ᶵ≀㉁ࡢホ౯ᩘࡇࡢ⤖ᯝࡣࠊ㑇ఏẘᛶ≀㉁ࡢᐃ⩏ࢆࠕ࠸ࡎࢀ࠿ࡢ㑇ఏẘᛶヨ㦂࡛㝧ᛶ㸦ࡓࡔࡋࠊ࣐࢘ࢫࣜࣥࣇ࢛࣮࣐
TK ヨ㦂ࡢࡳ࡛㝧ᛶࡢሙྜࢆ㝖ࡃ㸧ࡢ≀㉁ࠖ࡜ࡋࠊ㠀㑇ఏẘᛶ≀㉁ࢆ௨ୗࡢ࠸ࡎࢀ࠿࡟ヱᙜࡍࡿ≀㉁࡜ᐃ
⩏ࡋ࡚ࡀࢇཎ≀㉁ࢆศ㢮ࡋࡓሙྜࡢ⤖ᯝ࡛࠶ࡿ 12)ࠋ䞉Ames ヨ㦂㸦᚟ᖐ✺↛ኚ␗ヨ㦂㸧ࡀ㝜ᛶ࡛ in vivo 㑇ఏẘᛶヨ㦂ࢹ࣮ࢱࡀ↓࠸ࠋ䞉Ames ヨ㦂㸦᚟ᖐ✺↛ኚ␗ヨ㦂㸧ࢹ࣮ࢱࡀᚓࡽࢀ࡞࠸ࡀࠊ௚ࡢ㑇ఏẘᛶヨ㦂࡛㝜ᛶ࡛࠶ࡿࠋ
䞉ᚓࡽࢀ࡚࠸ࡿࡍ࡭࡚ࡢ㑇ఏẘᛶヨ㦂࡛㝜ᛶ࡛࠶ࡿࠋ
ࡋ࠿ࡋ࡞ࡀࡽࠊ㑇ఏẘᛶࡢホ౯ࡣ Ames ヨ㦂ࡢࡳ࡛ホ౯ࡉࢀ࡚ࡣ࠸࡞࠸ࡇ࡜࠿ࡽࠊ㑇ఏẘᛶ≀㉁࠾ࡼࡧ
㠀㑇ఏẘᛶ≀㉁ࡢᐃ⩏ࢆ௨ୗࡢࡼ࠺࡟ኚ᭦ࡋ 2,12)ࠊ㑇ఏẘᛶ࡜ࡀࢇཎᛶࡢᩥ⊩ࢆ෌ㄪᰝࡋࠊ᭷ᶵ໬ྜ≀ࢆ෌ศ㢮ࡋࡓࠋ
<㑇ఏẘᛶ≀㉁>
䞉In vitro ヨ㦂ࢹ࣮ࢱࡀ㝧ᛶ࡛࠶ࡾࠊࡑࢀࡀ in vivo 㑇ఏẘᛶヨ㦂࡛☜ㄆࡉࢀ࡚࠸ࡿࠋ
䞉In vivo ヨ㦂ࢹ࣮ࢱࡀ฼⏝࡛ࡁ࡞࠸ሙྜ࡟ࡣࠊin vitro ヨ㦂⤖ᯝࡀ౑ࢃࢀࡿࠋ

<㠀㑇ఏẘᛶ≀㉁>䞉Ames ヨ㦂㸦᚟ᖐ✺↛ኚ␗ヨ㦂㸧࡛㝜ᛶ࡛࠶ࡾࠊᇵ㣴⣽⬊ࢆ⏝࠸ࡿヨ㦂㸦ᰁⰍయ␗ᖖヨ㦂/ᑠ᰾ヨ㦂/㑇ఏᏊ✺↛ኚ␗ヨ㦂/࣐࢘ࢫࣜࣥࣇ࢛࣮࣐ TK ヨ㦂࡟ࡘ࠸࡚ࡣ ICH ࡟ࡼࡿ⌧⾜ࡢᇶ‽ุ࡛᩿㸧࡛㝜ᛶࠊ࠶ࡿ࠸ࡣᶆ‽ⓗ in vivo ヨ㦂࡛㝜ᛶ㸦ྠࡌ࢚ࣥࢻ࣏࢖ࣥࢺࡢሙྜࠊin vivo ヨ㦂⤖ᯝࢆ᥇⏝ࡍࡿ㸧࡛࠶ࡿࠋࡑࡢ⤖ᯝࠊ෌ホ౯ࡉࢀࡓ᭷ᶵ໬ྜ≀ࡣ pH6.7 ἲ࡛ࡣ 85 ≀㉁ࠊpH7.0 ἲ࡛ࡣ 32 ≀㉁࡛࠶ࡾࠊࡑࢀࡽࡣ㑇ఏẘᛶࡀࢇཎ≀㉁ࠊ㠀㑇ఏẘᛶࡀࢇཎ≀㉁࠾ࡼࡧ㠀ࡀࢇཎ≀㉁࡟ศ㢮ࡉࢀࠊࡑࢀࡽࡢ SHE CTA ⤖ᯝࡣ⾲ 8 ࡟♧ࡋࡓࡼ࠺࡟ࠊ㠀㑇ఏẘᛶࡀࢇཎ≀㉁ࡢ᳨ฟ⋡ࡣࠊpH6.7 ἲ࡛ࡣ 56.5%ࠊpH7.0 ἲ࡛ࡣ 83.3%࡛࠶ࡗࡓ 2)ࠋ

15

21

⾲ 8

SHE CTA ࡟࠾ࡅࡿ㠀㑇ఏẘᛶࡀࢇཎ≀㉁ࡢ᳨ฟ⋡㑇ఏẘᛶࡀࢇཎ≀㉁㠀㑇ఏẘᛶࡀࢇཎ≀㉁㠀ࡀࢇཎ≀㉁

≀㉁ᩘ

᳨ฟ⋡
㸦%㸧

≀㉁ᩘ

᳨ฟ⋡
㸦%㸧

≀㉁ᩘ

᳨ฟ⋡
㸦%㸧

SHE CTA

㝧ᛶ

20

71.4

13

56.5

3

8.8

㸦pH6.7 ἲ㸧

㝜ᛶ

3

10.7

10

43.7

26

76.5

Equivocal

5

17.9

0

0.0

5

14.7

ィ

28

SHE CTA

㝧ᛶ

9

64.3

5

83.3

1

8.3

㸦pH7.0 ἲ㸧

㝜ᛶ

4

28.6

1

16.7

11

91.7

Equivocal

1

7.1

0

0.0

0

0.0

ィ

14

23

6

34

12

DRP31 ࡛ࡣࠊᩥ⊩ㄪᰝࡋࡓ໬Ꮫ≀㉁࡟ࡘ࠸࡚ࠊIARC ࡀሗ࿌ࡋ࡚࠸ࡿࣄࢺ࡟ᑐࡍࡿࡀࢇཎᛶࣜࢫࢡศ㢮ࢆグ㍕ࡋ࡚࠸ࡿࡀࠊࡑࡢศ㢮࡜ CTA ⤖ᯝࡢ┦㛵ᛶ࡟ࡘ࠸࡚ࡢヲ⣽ࡣグ㍕ࡉࢀ࡚࠸࡞࠸ࠋ
ࡑࡇ࡛ࠊᙜ㈨ᩱ⦅⧩ጤဨ఍࡛ࡣࠊDRP31 ࡢ Table 11㹼13 ࡟グ㍕ࡉࢀ࡚࠸ࡿ≀㉁ࡢ࠺ࡕࠊࣄࢺ࡟ᑐࡍࡿ
ࡀࢇཎᛶࣜࢫࢡศ㢮ࡢグ㍕ࡀ࠶ࡾࠊࡋ࠿ࡶ SHE CTA ࡢ⤖ᯝࡀ᫂ࡽ࠿࡞ 169 ≀㉁࡟ࡘ࠸࡚ࠊSHE CTA ⤖ᯝ࡜ࣄࢺࡀࢇཎᛶ࡜ࡢ┦㛵ࢆㄪᰝࡋࡓࠋ࡞࠾ࠊIARC ࡢศ㢮࡟ࡘ࠸࡚ࡣࠊBenigni ࡽ 13) ࡢሗ࿌ࡶཧ⪃࡟ࡋࡓࠋࡑࡢ⤖ᯝࠊࢢ࣮ࣝࣉ 1㸦ࣄࢺ࡟ᑐࡋ࡚ࡀࢇཎᛶࡀ࠶ࡿ㸧࠾ࡼࡧࢢ࣮ࣝࣉ 2A㸦ࣄࢺ࡟ᑐࡋ࡚࠾ࡑࡽࡃ
ࡀࢇཎᛶࡀ࠶ࡿ㸧ࢆࣄࢺࡀࢇཎ≀㉁࡜ࡋࡓሙྜࠊSHE CTA㸦pH6.7 ἲࡲࡓࡣ pH7.0 ἲࡢ࠸ࡎࢀ࠿୍᪉࡛ࡶ
㝧ᛶࡢሙྜࢆ SHE CTA 㝧ᛶ࡜ุᐃ㸧࡟࠾ࡅࡿ୍⮴⋡ࡣ 55.0%ࠊឤᗘࡣ 93.7%ࠊ≉␗ᗘࡣ 32.1%࡜ࣄࢺࡀࢇཎ≀㉁ࡢ᳨ฟឤᗘࡢ㧗࠸⤖ᯝ࡛࠶ࡗࡓ㸦Appendix II㸧ࠋࡉࡽ࡟ࠊIARC ศ㢮ẖࠊ SHE CTA ࠾ࡼࡧࡀࢇཎᛶヨ㦂⤖ᯝࡢヨ㦂ἲẖ࡟ࡑࢀࡽࡢ┦㛵ࢆ⾲ 9-1 ࡟♧ࡋ
ࡓࠋ

16

22

⾲ 9-1 ࣄࢺ࡟࠾ࡅࡿࡀࢇཎᛶࣜࢫࢡศ㢮㸦IARC ศ㢮㸧࡜ SHE CTA ࠾ࡼࡧࡀࢇཎᛶヨ㦂⤖ᯝ࡜ࡢ┦㛵ᛶ
࢔ࢵࢭ࢖⣔

ุᐃ

ࣄࢺ࡟ᑐࡍࡿࡀࢇཎᛶ
ࢢ࣮ࣝࣉ 1ࢢ࣮ࣝࣉ 2A

ࢢ࣮ࣝࣉ 2B

ࢢ࣮ࣝࣉ 3

SHE CTA㝧ᛶ

6 (66.7%)

6 (100.0%)

12 (75.0%)

8 (36.4%)(pH 6.7 ἲ)㝜ᛶ

3 (33.3%)

0 (0.0%)4 (25.0%)

14 (63.8%)

ィ

9

6

16

22

SHE CTA㝧ᛶ

43 (95.6%)

13 (92.9%)

40 (87.0%)

24 (51.1%)(pH 7.0 ἲ)㝜ᛶ

2 (4.4%)1 (7.1%)6 (13.0%)23 (48.9%)

ィ

45

14

46

47

ᐇ㦂ື≀

㝧ᛶ

48 (100.0)

16 (94.1%)

50 (94.3%)

22 (39.3%)

࡟࠾ࡅࡿ

㝜ᛶ

0 (0.0)1 (5.9%)3 (5.7%)34 (60.7%)

ࡀࢇཎᛶヨ㦂

ィ

48

17

53

56

ࡇࢀࡽ≀㉁࡟ࡣࠊࡆࡗṑ㢮ࡢࡀࢇཎᛶヨ㦂࡛㝧ᛶࡢ 136 ≀㉁㸦᭷ᶵ໬ྜ≀㸸94 ≀㉁ࠊ↓ᶵ໬ྜ≀㸸42≀㉁㸧࡜㝜ᛶࡢ 38 ≀㉁㸦᭷ᶵ໬ྜ≀㸸34 ≀㉁ࠊ↓ᶵ໬ྜ≀㸸4 ≀㉁㸧ࡀྵࡲࢀ࡚࠸ࡿࡀࠊࡉࡽ࡟᭷ᶵ໬
ྜ≀࡜↓ᶵ໬ྜ≀࡟ศ㢮ࡋ࡚⾲ 9-2 ࡟♧ࡋࡓࠋ

1723

⾲ 9-2 ࣄࢺ࡟࠾ࡅࡿࡀࢇཎᛶࣜࢫࢡศ㢮㸦IARC ศ㢮㸧࡜ SHE CTA ࠾ࡼࡧࡀࢇཎᛶヨ㦂⤖ᯝ࡜ࡢ┦㛵ᛶ㸦᭷ᶵ໬ྜ≀㸧࢔ࢵࢭ࢖⣔ุᐃࣄࢺ࡟ᑐࡍࡿࡀࢇཎᛶ
ࢢ࣮ࣝࣉ 1ࢢ࣮ࣝࣉ 2A

ࢢ࣮ࣝࣉ 2B

ࢢ࣮ࣝࣉ 3

SHE CTA㝧ᛶ

3 (50.0%)

6 (100.0%)

10 (76.9%)

7 (33.3%)(pH 6.7 ἲ)㝜ᛶ

3 (50.0%)0 (0.0%)3 (23.1%)

14 (66.7%)

ィ

6

6

13

21

SHE CTA㝧ᛶ

11 (84.6%)

12 (92.3%)

34 (87.2%)

21 (50.0%)(pH 7.0 ἲ)㝜ᛶ

2 (15.4%)

1 (7.7%)5 (12.8%)

21 (50.0%)

ィ

13

13

39

42

ᐇ㦂ື≀

㝧ᛶ

15 (100.0)

15 (93.8%)

43 (93.5%)

21 (41.2%)

࡟࠾ࡅࡿ

㝜ᛶ

0 (0.0)1 (6.2%)3 (6.5%)30 (58.8%)

ࡀࢇཎᛶヨ㦂

ィ

15

16

46

51

㸦↓ᶵ໬ྜ≀㸧࢔ࢵࢭ࢖⣔ุᐃࣄࢺ࡟ᑐࡍࡿࡀࢇཎᛶ
ࢢ࣮ࣝࣉ 1ࢢ࣮ࣝࣉ 2A

ࢢ࣮ࣝࣉ 2B

ࢢ࣮ࣝࣉ 3

SHE CTA㝧ᛶ

3 (100.0%)

0 (0.0%)2 (66.7%)

1 (100.0%)(pH 6.7 ἲ)㝜ᛶ

0 (0.0%)0 (0.0%)1 (33.3%)

0 (0.0%)

ィ

3

0

3

1

SHE CTA㝧ᛶ

32(100.0%)

1 (100.0%)

6 (85.7%)

3 (60.0%)(pH 7.0 ἲ)㝜ᛶ

0 (0.0%)0 (0.0%)1 (14.3%)

2 (40.0%)

ィ

32

1

7

5

ᐇ㦂ື≀

㝧ᛶ

33 (100.0)

1 (100.0%)

7 (100.0%)

1 (20.0%)

࡟࠾ࡅࡿ

㝜ᛶ

0 (0.0)0 (0.0%)0 (0.0%)4 (80.0%)ࡀࢇཎᛶヨ㦂

ィ

33

1

7

5

ࢢ࣮ࣝࣉ 1 ࡟࠾ࡅࡿ≀㉁࡛ࡢ SHE CTA ࡢ᭷ᶵ໬ྜ≀ࡢ㝧ᛶ⋡ࡣࠊpH 6.7 ἲ࡛ࡣ 50.0%ࠊpH 7.0 ἲ࡛ࡣ
84.6%࡛࠶ࡾࠊᐇ㦂ື≀࡟࠾ࡅࡿࡀࢇཎᛶヨ㦂ࡢ⤖ᯝ㸦100.0%㸧࡜ẚ㍑ࡍࡿ࡜౛ᩘࡀᑡ࡞࠸ࡶࡢࡢప࠸್
࡛࠶ࡗࡓࠋ୍᪉ࠊ↓ᶵ໬ྜ≀࡟ࡘ࠸࡚ࡣࠊSHE CTA ࠾ࡼࡧࡀࢇཎᛶヨ㦂࡜ࡶ࡟ 100.0%ࡢ㝧ᛶ⋡࡛࠶ࡗࡓࠋ
ࢢ࣮ࣝࣉ 2A ࡟࠾ࡅࡿ᭷ᶵ໬ྜ≀ࡢ㝧ᛶ⋡ࡣࠊpH 6.7 ἲ࡛ࡣ 100.0%ࠊpH 7.0 ἲ࡛ࡣ 92.3%࡛࠶ࡾࠊࡀࢇ
ཎᛶヨ㦂ࡢ⤖ᯝ㸦93.8%㸧࡜ྠ➼࡛࠶ࡗࡓࠋ↓ᶵ໬ྜ≀࡟ࡘ࠸࡚ࡣ 1 ≀㉁ࡢࡳ࡛࠶ࡿࡀࠊ࠸ࡎࢀࡢ pH ࡛ࡶ㝧ᛶ࡛࠶ࡗࡓࠋ
ࢢ࣮ࣝࣉ 2B㸦ࣄࢺ࡟ᑐࡋ࡚ࡀࢇཎᛶࡢྍ⬟ᛶࡀ࠶ࡿ㸧࡟࠾ࡅࡿ᭷ᶵ໬ྜ≀ࡢ SHE CTA ࡢ㝧ᛶ⋡㸦76.9
18

24

࠾ࡼࡧ 87.2%㸧ࡣࠊࡀࢇཎᛶヨ㦂ࡢ⤖ᯝ㸦93.5%㸧ࡼࡾࢃࡎ࠿࡟పୗࡋࡓࠋ↓ᶵ໬ྜ≀ࡶྠᵝ࡟ࠊSHE CTAࡢ㝧ᛶ⋡㸦66.7 ࠾ࡼࡧ 85.7%㸧ࡣࡀࢇཎᛶヨ㦂ࡢ⤖ᯝ㸦100.0%㸧ࡼࡾపୗࡋࡓࠋ
ࢢ࣮ࣝࣉ 3㸦ࣄࢺ࡟ᑐࡍࡿⓎ⒴ᛶࡀศ㢮࡛ࡁ࡞࠸㸧࡟࠾ࡅࡿ᭷ᶵ໬ྜ≀ࡢ SHE CTA ࡢ㝧ᛶ⋡㸦ഇ㝧ᛶ⋡࡜ྠࡌ㸧ࡣࠊpH 6.7 ἲ࡛ࡣ 33.3%ࠊpH 7.0 ἲ࡛ࡣ 50.0%࡛࠶ࡾࠊࡀࢇཎᛶヨ㦂ࡢ⤖ᯝ㸦41.2%㸧࡜࡯ࡰྠ➼࡛࠶ࡗࡓࠋࡑࡢ㝜ᛶ⋡㸦≉␗ᗘ࡜ྠࡌ㸧ࡶࠊpH 6.7 ἲ࡛ࡣ 66.7%ࠊpH 7.0 ἲ࡛ࡣ 50.0%࡛ࠊࡀࢇཎᛶヨ㦂㸦58.6%㸧࡜࡯ࡰྠ➼࡛࠶ࡗࡓࠋ

8.

ヨ㦂᪉ἲࡢಙ㢗ᛶ1) ᪋タ㛫ኚື(1) ECVAM ࡢࣉࣞࣂࣜࢹ࣮ࢩࣙࣥ◊✲SHE CTA ࡢ᪋タ㛫ࡢ෌⌧ᛶࡣࠊpH6.7 ࠾ࡼࡧ pH7.0 ἲࡢ࡝ࡕࡽࡶࠊ⾲ 10 ࡟♧ࡍࡼ࠺࡟ 4 ࡘࡢࡀࢇཎ≀㉁࡜ 2 ࡘࡢ㠀ࡀࢇཎ≀㉁ࠊィ 6 ≀㉁ࢆ⏝࠸᳨࡚ドࡉࢀࡓ㸦Benzo[a]pyrene ࡣ㝧ᛶᑐ↷≀㉁࡜ࡋ࡚ࡢホ౯㸧6,7)ࠋ

⾲ 10 ECVAM ࣉࣞࣂࣜࢹ࣮ࢩࣙࣥ◊✲᳨࡛ウࡉࢀࡓ⿕㦂≀㉁໬Ꮫ≀㉁CAS ␒ྕ

ࡆࡗṑ㢮ࡀࢇཎᛶ㸦ฟ඾㸧

Benzo[a]pyrene

50-32-8

࠶ࡾ㸦IARC, 2009㸧

Anthracene120-12-7

࡞ࡋ㸦IARC, 2009㸧

2,4-Diaminotoluene

95-80-7

࠶ࡾ㸦IARC, 2009㸧

3-Methylcholanthrene

56-49-5

࠶ࡾ㸦Gold and Zeiger, 1997㸧o-Toluidine HCl

636-21-5

࠶ࡾ㸦NTP㸧

Phthalic anhydride

85-44-9࡞ࡋ㸦NTP㸧

pH7.0 ἲ࡟ࡘ࠸࡚ࡣࠊ4 ᪋タ࡟࡚ヨ㦂ࢆ⾜࠸ࠊホ౯ࡋࡓ⤖ᯝࠊPhthalic anhydride ࢆ㝖ࡃ඲࡚ࡢ⿕㦂≀㉁࡟
ࡘ࠸ุ࡚ᐃ⤖ᯝࡀ୍⮴ࡋࠊື≀ࡀࢇཎᛶࡢ⤖ᯝ࡜ࡶྜ⮴ࡋࡓࠋPhthalic anhydride ࡟ࡘ࠸࡚ࡣࠊ1 ᪋タ࡟࡚
1 ⃰ᗘࡢࡳᙧ㉁㌿᥮ࢥࣟࢽ࣮ࡢ᭷ព࡞ቑຍࡀ࠶ࡾࠊഴྥ᳨ᐃ࡛ࡶ᭷ព࡞ഴྥࢆ♧ࡋࡓࡇ࡜࠿ࡽ㝧ᛶ࡜ࡢุᐃ࡟࡞ࡗࡓࡀࠊ௚ࡢ᪋タ࡛ࡣື≀ࡀࢇཎᛶࡢホ౯⤖ᯝ࡜ྠࡌࡃࠊ㝜ᛶ࡜ุᐃࡉࢀࡓࠋ
pH6.7 ἲ࡟ࡘ࠸࡚ࡣࠊ3 ᪋タ࡟࡚ヨ㦂ࢆ⾜࠸ࠊホ౯ࡋࡓ⤖ᯝࠊ඲࡚ࡢ⿕㦂≀㉁ࡢุᐃ⤖ᯝࡀ୍⮴ࡋࠊPhthalicanhydride ࢆ㝖࠸࡚ࡣື≀ࡀࢇཎᛶヨ㦂⤖ᯝ࡜ࡶྜ⮴ࡋࡓࠋPhthalic anhydride ࡢ SHE CTA ࡢホ౯⤖ᯝࡣྛ᪋タ࡜ࡶ㝧ᛶ࡜ุᐃࡋࡓࡀࠊື≀ࡀࢇཎᛶࢆ♧ࡉ࡞࠸࡜ࡍࡿ NTP ࡢሗ࿌࡜▩┪ࡋࠊࡲࡓୖグࡢ pH7.0 ἲࡢ SHE CTA ࡟࠾ࡅࡿホ౯⤖ᯝ࡜ࡶ␗࡞ࡿࠋࡇࡢ⌮⏤࡜ࡋ࡚ Phthalic anhydride ࡢ␗࡞ࡿ pH ࡛ࡢᏳᐃᛶࡢ
㐪࠸ࡀᣦ᦬ࡉࢀ࡚࠸ࡿࠋ
ࡘࡲࡾ Phthalic anhydride ࡣỈ⎔ቃୗ࡛㏿ࡸ࠿࡟ຍỈศゎࡋࠊ㠀㑇ఏẘᛶࡢ Phthalicacid ࡟ኚ໬ࡍࡿࡀࠊSHE CTA ࡛ࡣ౑⏝ࡍࡿ⦆⾪ᾮࡀ␗࡞ࡾࠊࡼࡾప࠸ pH ࡛ࡣ Phthalic anhydride ࡢ༙ῶᮇࡀ㛗ࡃ࡞ࡿࡇ࡜࠿ࡽࠊ␗࡞ࡗࡓヨ㦂⤖ᯝࡀᚓࡽࢀࡓྍ⬟ᛶࡀ♧ࡉࢀ࡚࠸ࡿࠋ ୖグࡢ᪋タ㛫ࡢヨ㦂⤖ᯝࢆ⥲ྜⓗ࡟ホ౯ࡋࡓ⤖ᯝࠊྛ᪋タ࡛ࡢ SHE CTA ࡢホ౯⤖ᯝࡣᇶᮏⓗ࡟୍⮴ࡍ1925ࡿࡶࡢ࡛࠶ࡾࠊpH6.7 ἲ࠾ࡼࡧ pH7.0 ἲ࡜ࡶࡑࡢ᪋タ㛫෌⌧ᛶࡣ㧗࠸࡜ุ᩿ࡉࢀࡓࠋ
ุᐃ⤖ᯝ࡟㛵ࡋ࡚ࠊࢥࣟࢽ࣮ࡢ࢝࢘ࣥࢺࡀ୺ほⓗ࡛࠶ࡿࡇ࡜࡟ࡼࡿุᐃࡢࡤࡽࡘࡁࡀᠱᛕࡉࢀ࡚࠸ࡓ
ࡀࠊ෗┿ᶆᮏࡢᥦ౪ࡀุᐃࡢࡤࡽࡘࡁࡢᢚไ࡟኱ࡁࡃᐤ୚ࡋ࡚࠸ࡿ࡜⪃࠼ࡽࢀࡓࠋ
(2) ࡑࡢ௚ࡢᩥ⊩ሗ࿌࡜ࡢ୍⮴ᛶ
pH7.0 ἲ࡟ࡘ࠸࡚ࡣࠊ㢮ఝࣉࣟࢺࢥࣝ࡟ࡼࡿ෌⌧ᛶࢆホ౯ࡋࡓᩥ⊩ሗ࿌ 14)࡞ࡽࡧ࡟ 2 ᶵ㛵௨ୖ࡛ホ౯ࡉࢀࡓ≀㉁ࡢ 87.7㸣㸦57/65㸧ࡀ୍⮴ࡍࡿ࡜ࡋࡓ DRP31 ࢆཧ↷ࡋࡓࠋࡲࡓ pH6.7 ἲ࡟ࡘ࠸࡚ࡣࠊ㢮ఝࣉࣟࢺࢥࣝ࡟ࡼࡿ෌⌧ᛶࢆホ౯ࡋࡓᩥ⊩ሗ࿌ 15,16)࠾ࡼࡧୖグ DRP31 ࢆཧ↷ࡋࡓࠋࡇࢀࡽࡢሗ࿌࠿ࡽࠊⰋዲ࡞᪋タ㛫෌⌧ᛶࢆ♧ࡍ࡜⪃࠼ࡽࢀࡓࠋ

2)

᪋タෆኚື
᪋タෆ෌⌧ᛶ࡟ࡘ࠸࡚ࡣࠊpH6.7 ἲ࠾ࡼࡧ pH7.0 ἲࡢ୧᪉࡟ࡘ࠸࡚ࠊࣉࣞࣂࣜࢹ࣮ࢩࣙࣥ◊✲ࡀ⾜ࢃࢀホ౯ࡉࢀ࡚࠸ࡿࡀࠊ㝧ᛶᑐ↷࡜ࡋ࡚౑⏝ࡋࡓ 1 ໬ྜ≀㸦Benzo[a]pyrene㸧ࡢ⤖ᯝࡢࡳࡢホ౯࡜࡞ࡗ࡚࠸ࡿ 6,7)ࠋ
pH6.7 ἲ࡛ࡣࠊࢥ࣮ࢻ໬ࡏࡎ࡟ᐇ᪋ࡋࡓ⤖ᯝ࡜ࢥ࣮ࢻ໬ࡋ࡚ᐇ᪋ࡋࡓ 3 ᪋タࡢ⤖ᯝࢆẚ㍑ࡋ࡚ホ౯ࡋ࡚࠸ࡿࠋࡑࡢ⤖ᯝࠊ1 ᪋タ࡛ࢥࣥࢺ࣮ࣟࣝࡢᙧ㉁㌿᥮⋡ࡀ௚ࡢ 2 ᪋タ࡜␗࡞ࡿ್ࡀᚓࡽࢀ࡚࠸ࡿࡀࠊ㝧ᛶᑐ↷≀㉁࡛࠶ࡿ Benzo[a]pyrene ࡢ⤖ᯝࡣ࡯ࡰྠᵝࡢ್ࡀᚓࡽࢀ࡚࠸ࡿࠋ୍᪉ࠊpH7.0 ἲ࡛ࡣࢥ࣮ࢻ໬ࡋࡓ
Benzo[a]pyrene ࢆ⏝࠸࡚ 1 ᪋タࡢࡳࡀ 3 ᅇࡢ⧞ࡾ㏉ࡋᐇ㦂ࢆ⾜࠸ࠊⰋዲ࡞⤖ᯝࡀᚓࡽࢀ࡚࠸ࡿࠋ
ESAC ࡢሗ࿌ 10)࡟࠶ࡿࡼ࠺࡟ࠊࣉࣞࣂࣜࢹ࣮ࢩࣙࣥ◊✲࡛ࡣ᪋タෆ෌⌧ᛶࢆㄪ࡭ࡿࡓࡵࡢ㐺ษ࡞ᐇ㦂ࢹࢨ࢖ࣥࡀ⤌ࡲࢀ࡚࠾ࡽࡎࠊ⌧ᅾࡢホ౯ᇶ‽࡛⪃࠼ࡓሙྜࠊ᪋タෆ෌⌧ᛶ࡟ࡘ࠸࡚ࡣ༑ศ࡞ホ౯ࡀ⾜ࢃࢀ࡚࠸ࡿ࡜ࡣ⤖ㄽ࡛ࡁ࡞࠸ࠋ

9.

ヨ㦂᪉ἲࡢࢹ࣮ࢱࡢ㉁䞉ࣉࣞࣂࣜࢹ࣮ࢩࣙࣥሗ࿌᭩࡟ࡼࡿ࡜ࠊࣉࣞࣂࣜࢹ࣮ࢩࣙࣥ◊✲ࡢ᪋タෆ෌⌧ᛶࡢ☜ㄆࢆ」ᩘࡢ໬ྜ
≀࡛ᐇ᪋ࡋ࡞࠿ࡗࡓ࡞࡝᪉ἲ࡟୙༑ศ࡞Ⅼࡀ࠶ࡗࡓࡶࡢࡢࠊヨ㦂ᐇ᪋࡟ᚲせ࡞ࢫ࢟ࣝࡢカ⦎ࢆཷࡅࡓ᪋タ࡛ࡢ᪋タෆ/᪋タ㛫෌⌧ᛶࡣ☜ಖࡉࢀ࡚࠾ࡾࠊࢹ࣮ࢱࡢ㉁ࡣᢸಖࡉࢀ࡚࠸ࡿ࡜⪃࠼ࡽࢀࡿࠋ䞉ホ౯ୖ᭱ࡶ㔜せ࡞ࢹ࣮ࢱࡣࠊᙧ㉁㌿᥮ࢥࣟࢽ࣮ࡢィ್࡛࠶ࡿࠋ⏬ീ࢔ࢺࣛࢫ࡟ࡼࡾᶆ‽໬ࡉࢀ࡚࠸ࡿࡶࡢࡢࠊᙧ㉁㌿᥮ࢥࣟࢽ࣮ࡣ┠どุู࡛ࡍࡿࡓࡵࠊุูἲࡢᐈほᛶ࡟ஈࡋ࠸ࠋࡇࡢࡓࡵࠊࢹ࣮
ࢱࡢ㉁ࡢᢸಖ࡟ࡣࠊࢥ࣮ࢻ໬ࡋ࡚ྲྀᚓࡉࢀࡓࢹ࣮ࢱ࡛࠶ࡿࡇ࡜ࡀ㔜せ࡜࡞ࡿࠋࣉࣞࣂࣜࢹ࣮ࢩࣙࣥ
◊✲ࡢ໬ྜ≀ࡣࢥ࣮ࢻ໬ࡉࢀࠊᙧ㉁㌿᥮ࢥࣟࢽ࣮ࡢィࡣࢥ࣮ࢻ໬ࡋ࡚⾜ࢃࢀࡓࠋᚑࡗ࡚ࠊྲྀᚓࡉ

ࢀࡓࢹ࣮ࢱࡢ㉁ࡣᢸಖࡉࢀ࡚࠸ࡿ࡜⪃࠼ࡽࢀࡿࠋ
䞉ヨ㦂ࡢጇᙜᛶࢆᢸಖࡍࡿࡓࡵࡢࡶ࠺୍ࡘࡢ㔜せ࡞ DRP31 ࡛᥇⏝ࡉࢀࡓࢹ࣮ࢱࡣࢥ࣮ࢻ໬ࡋ࡚ྲྀᚓ
ࡉࢀࡓࡶࡢ࡛ࡣ࡞࠸ࠋࡋ࠿ࡋ࡞ࡀࡽࣉࣞࣂࣜࢹ࣮ࢩࣙࣥ◊✲ࡢุᐃᇶ‽ࢆ௨࡚ DRP31 ࢹ࣮ࢱࢆ෌ホ౯ࡋࡓ⤖ᯝࠊឤᗘࡸ≉␗ᗘ࡟኱ᕪࡀ࡞࠿ࡗࡓࡇ࡜࠿ࡽࠊࡇࢀࡽ࡟ࡘ࠸࡚ࡶࢹ࣮ࢱࡢ㉁ࡣᢸಖࡉࢀ࡚
࠸ࡿ࡜⪃࠼ࡽࢀࡿࠋ

20

26

10.

ヨ㦂᪉ἲ࡟㛵ࡍࡿ௚ࡢ⛉Ꮫⓗ࡞ሗ࿌

SHE CTA ࡜ྠᵝ࡟⣽⬊ࡢ CTA ࡟ࡣ௨ୗࡢ᪉ἲࡀ㛤Ⓨࡉࢀ࡚࠸ࡿࠋ
1) ⣽⬊ᰴ BALB/c3T3 ࢆ⏝࠸ࡿ CTA2) ⣽⬊ᰴ Bhas42 ࢆ⏝࠸ࡿ CTAͤ3) ⣽⬊ᰴ C3H10T1/2 ࢆ⏝࠸ࡿ CTA
4) JB6 ⣽⬊ࢆ⏝࠸ࡿ CTA
2) Bhas42 ࢆ⏝࠸ࡿ CTA ࡀ 2016 ᖺ 1 ᭶ OECD ࢞࢖ࢲࣥࢫ 17)࡜࡞ࡗࡓࠋͤୖグࡢ 1) ࠥ3) ࡟ࡘ࠸࡚ࡣࠊSHE CTA ࡜ྠᵝ࡟ᙧ㉁㌿᥮ࡋࡓ⣽⬊࡟ࡘ࠸࡚ࡢホ౯࡛࠶ࡿࡀࠊᙧᡂࡉࢀ
ࡓࢥࣟࢽ࣮୰ࡢᙧ㉁㌿᥮ࡋࡓࢥࣟࢽ࣮ᩘࢆᩘ࠼ࡿࡢ࡛ࡣ࡞ࡃࠊ༢ᒙࡢṇᖖ⣽⬊୰࡛ቑṪࡋࡓᙧ㉁㌿᥮⣽⬊ࡀᙧᡂࡍࡿࣇ࢛࣮࢝ࢫ㸦ᙧ㉁㌿᥮⣽⬊ࡢ㞟ྜయ㸧ࡢᩘࢆィࡍࡿࡶࡢ࡛࠶ࡿࠋ4㸧ࡢ JB6 ⣽⬊࡟ࡘ࠸࡚ࡣࠊ㌾ᐮኳᇵᆅ࡛ࡢቑṪ⬟㸦㊊ሙ౫Ꮡᛶࡢ႙ኻ㸧ࢆᣦᶆ࡜ࡍࡿヨ㦂࡛࠶ࡿࠋ

11.

3Rs ࡬ࡢ㛵୚㸦ື≀⚟♴㠃࠿ࡽࡢጇᙜᛶ㸧SHE CTA ࡛ࡣࢩࣜ࢔࣭ࣥࣁ࣒ࢫࢱ࣮ࡢ⫾௘࠿ࡽᚓࡓึ௦ᇵ㣴⣽⬊ࢆ⏝࠸ࡿࡓࡵࠊື≀ࢆ≛≅࡟ࡍࡿᚲせࡀ࠶ࡿࠋࡑࡢࡓࡵࠊ௚ࡢᇵ㣴⣽⬊㸦Bhas42㸪BALB/c 3T3㸧ࢆ⏝࠸ࡿ CTA ࡜ẚ㍑ࡋ࡚ࠊື≀⚟♴㠃࡛ࡣ୙฼࡛࠶ࡿࡇ࡜ࡣྰࡵ࡞࠸ࠋࡋ࠿ࡋ࡞ࡀࡽࠊ୍༉ࡢዷፎື≀⏤᮶ࡢ⫾௘⣽⬊࡛⣙ 50 ᅇࡢ CTA ࡀᐇ᪋ྍ⬟࡛࠶ࡿࡇ࡜ࠊ࠾ࡼࡧᮏヨ㦂ࢆྵࡴ CTA ࡣࡀࢇ໬㐣⛬࡟࠾ࡅࡿ⏕≀Ꮫⓗࣉࣟࢭࢫࢆホ౯ࡍࡿࡓࡵࠊ㐺ษ࡟ᐇ᪋ࡍࡿࡇ࡜࡟ࡼࡗ࡚ࠊከᩘࡢື≀ࢆ౑⏝ࡍࡿࡀࢇཎᛶヨ㦂࠾ࡼࡧ㛵㐃ヨ㦂ࢆᅇ㑊ࠊ࠶ࡿ࠸ࡣ┬␎ࡍࡿࡇ࡜ࡀᮇᚅ࡛ࡁࡿࡓࡵࠊື≀⚟♴࡬ࡢᐤ୚ࡣ኱ࡁ࠸࡜⪃࠼ࡽࢀࡿࠋ

12. ヨ㦂᪉ἲࡢ᭷⏝ᛶ࡜㝈⏺
1)᭷⏝ᛶ(1) CTA䞉ࡀࢇ໬ࡢ㐣⛬ࢆㄝ᫂ࡍࡿከẁ㝵ㄝ࡟ࡼࢀࡤࠊኚ␗ཎ≀㉁࡟ࡼࡾ࢖ࢽࢩ࢚࣮ࢩࣙࣥస⏝ࢆཷࡅࡓ⣽⬊ࡣࠊ᭦࡟ࡑࡢᚋࡉࡲࡊࡲ࡞せᅉ࡟ࡼࡗ࡚ᝏᛶᙧ㉁ࢆ⋓ᚓࡍࡿ࡜⪃࠼ࡽࢀ࡚࠸ࡿࠋCTA ࡣࠊᇵ㣴ṇᖖ⣽⬊ᡈ࠸ࡣᰴ⣽⬊ࡀᙧែᙧ㉁㌿᥮ࡍࡿࡇ࡜ࢆᣦᶆ࡟ࡋ࡚ࠊ໬Ꮫ≀㉁ࡢᙧ㉁㌿᥮⬟ࡢ᭷↓ࢆணࡍࡿ

in vitro ヨ㦂ἲࡢ୍ࡘ࡜ࡋ࡚ࡢ≉ᚩࡀ࠶ࡿࠋ
䞉௚ࡢࡀࢇཎᛶண

ヨ㦂⤖ᯝ࡜࡜ࡶ࡟ࠊWoE ࡢ୍ࡘ࡜ࡋ࡚ CTA ࡢ⤖ᯝࢆ฼⏝ࡍࡿࡇ࡜ࡀ࡛ࡁࡿࠋᮏ

ヨ㦂ἲࡣ⌧⾜ࡢࡀࢇཎᛶヨ㦂࡟᏶඲࡟⨨ࡁ᥮ࢃࡿヨ㦂ἲ࡜ࡋ࡚ࡢ఩⨨࡙ࡅ࡛ࡣ࡞ࡃࠊ⿵ຓⓗ࡞ヨ㦂㸦౛࠼ࡤࠊࡀࢇཎᛶヨ㦂ࡢᐇ᪋㡰఩ࢆỴࡵࡿࡓࡵࡢࢫࢡ࣮ࣜࢽࣥࢢࠊࡀࢇཎᛶヨ㦂ࡀせồࡉࢀ࡞࠸࢝ࢸࢦ࣮ࣜࡢ་⸆ရࡢࡀࢇཎᛶ㛵㐃᝟ሗࡢྲྀᚓ࡞࡝㸧࡜ࡋ࡚఩⨨࡙ࡅࡿࡇ࡜ࡣྍ⬟࡛࠶ࡿࠋ䞉3Rs ࡢほⅬ࠿ࡽࡶ᭷⏝࡛࠶ࡿࠋ

21

27

(2) SHE CTACTA ࡢ᭷⏝ᛶ࡟ຍ࠼࡚௨ୗࡢ᭷⏝ᛶࢆㄆࡵࡿࠋ
䞉DRP31 ࡢ 245 ໬Ꮫ≀㉁࡛ホ౯ࡉࢀࡓ⤖ᯝ࡟࠾࠸࡚ࠊSHE CTA ࡢ⤖ᯝ࡜ࡀࢇཎᛶヨ㦂⤖ᯝ࡜ࡢ┦㛵ࡀㄆࡵࡽࢀࡿࠋ
䞉SHE CTA ࡢ⣽⬊ㄪ〇ࡣ෾⤖ಖᏑࡋࡓึ௦஧ಸయ⣽⬊ࢆ⏝࠸ࡿࡀࠊᚲせ࡜ࡍࡿື≀ᩘࡣࡀࢇཎᛶヨ㦂࡟⏝࠸ࡿື≀ᩘ࡟ẚ࡭ࡣࡿ࠿࡟ᑡ࡞࠸ࠋ
䞉3Rs ࡢほⅬ࠿ࡽ᭷⏝࡛࠶ࡿࡇ࡜࡟ຍ࠼࡚ࠊ୙Ṛ໬ᇵ㣴⣽⬊ࢆ⏝࠸ࡿ CTA ࡜ẚ㍑ࡋ࡚ࠊึ௦ᇵ㣴⣽⬊ࢆ⏝࠸ࡿࡇ࡜࠿ࡽࠊࡼࡾṇᖖ⣽⬊࡟㏆࠸≧ែࢆ⥔ᣢࡋࡓᐇ㦂⣔࡛࠶ࡿ࡜⪃࠼ࡽࢀࡿࠋ

2)

ၥ㢟Ⅼ㸦㝈⏺ࠊ୙฼Ⅼࠊ㐺⏝㝈⏺㸧(1) CTA䞉CTA ࡛⤖ᯝࡀ㝧ᛶ࡛࠶ࡗ࡚ࡶࠊ໬Ꮫ≀㉁ࡢᙧ㉁㌿᥮⬟࠿ࡽࠊࡑࡢ໬Ꮫ≀㉁ࡢ in vivo ࡟࠾ࡅࡿࡀࢇཎ
ᛶࡢᙉࡉࠊ⮚ჾ≉␗ᛶࠊ࠾ࡼࡧ✀≉␗ᛶ࡟㛵ࡍࡿ᝟ሗࡣᚓࡽࢀ࡞࠸ࠋ
䞉CTA ࡢ࢚ࣥࢻ࣏࢖ࣥࢺ࡛࠶ࡿᙧ㉁㌿᥮ࡢⓎ⏕ᶵᗎࡀ᫂ࡽ࠿࡟࡞ࡗ࡚࠸࡞࠸ࡓࡵࠊ㠀㑇ఏẘᛶࡀࢇཎ≀㉁ࡢస⏝ᶵᗎ࡟㛵ࡍࡿ᝟ሗࡣᚓࡽࢀ࡞࠸ࠋ
䞉୍⯡ⓗ࡞ in vitro ヨ㦂࡜ྠᵝ࡟ࠊ⤖ᯝࡢጇᙜᛶࡣᶆ‽໬ࡉࢀࡓࣉࣟࢺࢥࣝ࡟ᚑࡗ୍࡚㐃ࡢヨ㦂᧯సࡀṇ☜࡟⾜ࢃࢀࡿࡇ࡜࡟ࡼࡗ࡚ಖドࡉࢀࡿࠋCTA ࡢ⤖ᯝࡢṇ☜ᛶࢆಖドࡍࡿࡓࡵࠊヨ㦂᧯సࡣࡶࡕࢁ
ࢇࠊᙧ㉁㌿᥮ࢥࣟࢽ࣮/ࣇ࢛࣮࢝ࢫࡢุูࢫ࢟ࣝࡢ⩦⇍ࡀ㔜せ࡛࠶ࡿࠋᶆ‽≀㉁࡛ㄏᑟࡉࢀࡓᙧ㉁㌿᥮ࢥࣟࢽ࣮/ࣇ࢛࣮࢝ࢫࡢ⏬ീ࢔ࢺࣛࢫ➼ࢆ฼⏝ࡋࡓࢫ࢟ࣝ☜ㄆࡣ㔜せ࡛࠶ࡿࠋ䞉ᙧ㉁㌿᥮ࢥࣟࢽ࣮/ࣇ࢛࣮࢝ࢫࡢ⮬ືุูἲࡢ㛤Ⓨࡀ㐍ࢇ࡛࠸࡞࠸ࡓࡵࠊ⌧᫬Ⅼ࡛ࡢከ᳨య㧗㏿ࢫࢡ࣮ࣜࢽࣥࢢࡢሗ࿌ᩘࡣᑡ࡞࠸ࠋᗈ⠊࡞໬Ꮫ≀㉁ࡢᙧ㉁㌿᥮⬟ࢆ CTA ୍࡛ḟࢫࢡ࣮ࣜࢽࣥࢢࡍࡿࡓࡵ
࡟ࡣࠊヨ㦂⣔ࡢ᭦࡞ࡿᨵⰋࠊᡈ࠸ࡣࠊᙧ㉁㌿᥮ࢥࣟࢽ࣮/ࣇ࢛࣮࢝ࢫࡢ㠉᪂ⓗุู᪉ἲࡢ㛤Ⓨࡀᚲせ࡛࠶ࡿࠋ(2) SHE CTACTA ࡟࠾ࡅࡿၥ㢟Ⅼ࡟ຍ࠼࡚௨ୗࡢၥ㢟Ⅼࢆㄆࡵࡿࠋ
䞉SHE CTA ࡟ࡣࠊ࢖ࢽࢩ࢚࣮ࢩࣙࣥ࡜ࣉ࣮ࣟࣔࢩࣙࣥస⏝࡜ࢆ༊ูࡍࡿヨ㦂ἲࡀグ㍕ࡉࢀ࡚࠸࡞࠸ࡓ
ࡵࠊ࢖ࢽࢩ࢚࣮ࢩࣙࣥ/ࣉ࣮ࣟࣔࢩࣙࣥస⏝࡜࠸ࡗࡓほⅬ࡛ࡢ⿕㦂≀㉁ࡢస⏝ᶵᗎࢆ᫂ࡽ࠿࡟ࡍࡿࡇ
࡜ࡣ࡛ࡁ࡞࠸ࠋ
䞉SHE CTA ࡛ࡣࠊࣉࣞࣂࣜࢹ࣮ࢩࣙࣥ◊✲ࡀ⾜ࢃࢀࡓ 6 ≀㉁㸦ణࡋࠊ㑅ᢥࡉࢀࡓ≀㉁࡟ࡣࠊస⏝࣓࢝ࢽࢬ࣒࡟೫ࡾࡀ࠶ࡿ࡜ࡉࢀ࡚࠸ࡿ㸧࡟ࡘ࠸࡚ࡣࠊࡀࢇཎᛶヨ㦂⤖ᯝ࡜ࡢ┦㛵㸦ឤᗘࠊ≉␗ᗘࠊഇ㝧ᛶ࣭ഇ㝜ᛶࡢ㆑ูࡢ⤖ᯝ㸧ࡣ☜ㄆࡉࢀ࡚࠸ࡿࡀࠊ㑅ᢥࡉࢀࡓ≀㉁࡟ࡣࠊస⏝ᶵస࡟೫ࡾࡀ࠶ࡾ㸪໬Ꮫ≀㉁ࡢᩘࡸ✀㢮ࡶ༑ศ࡛ࡣ࡞࠸ࠋホ౯ࡉࢀࡓ໬Ꮫ≀㉁࡟ࡣ㑇ఏẘᛶࢆ᭷ࡍࡿࡀࢇཎ≀㉁ࡀከࡃྵ
ࡲࢀࠊ㠀㑇ఏẘᛶࡀࢇཎ≀㉁㸦͆┿ࡢ㸦bona fide㸧㠀㑇ఏẘᛶࡀࢇཎ≀㉁͇ࡢᐃ⩏࡟ࡣ㆟ㄽࡢవᆅࡀ
࠶ࡿ㸧ࡀᑡ࡞࠸ࡓࡵࠊ㠀㑇ఏẘᛶࡀࢇཎ≀㉁ࡢ᳨ฟ⣔࡜࠸࠺ほⅬ࡛ࡢ᭷⏝ᛶࡢ᳨ドࡣ୙༑ศ࡛࠶ࡿࠋ

22

28

3)

௒ᚋࡢㄢ㢟
⣽⬊ࡀᙧ㉁㌿᥮ࡍࡿస⏝ᶵᵓࡢゎ᫂ࡀ㔜せ࡛࠶ࡿࠋࡋ࠿ࡋࠊࡇࡢㄢ㢟ࡣࣁ࣮ࢻࣝࡀ㧗࠸ࠋ䞉ࣄࢺ⣽⬊࡛ࡶ㉳ࡇࡾ࠺ࡿ࠿ࢆ෌⌧ࡍࡿࡓࡵ࡟ࡣࠊࣄࢺ⣽⬊ࢆ⏝࠸ࡓ CTA ࡀ㛤Ⓨࡉࢀࡿࡇ࡜ࡀᮃࡲࡋ
࠸ࠋ㸦In vivo ࡟࠾ࡅࡿࠊື≀࠿ࡽࣄࢺ࡬ࡢእᤄ࡜ྠࡌၥ㢟ࢆྵࢇ࡛࠸ࡿࠋ㸧䞉ᙧែᙧ㉁㌿᥮ࢥࣟࢽ࣮ࡢุᐃࡢᐈほᛶࡢ☜ಖࡀᐇົୖᚲせ࡛࠶ࡿࠋ䞉┿ࡢ㸦bona fide㸧㠀㑇ఏẘᛶࡀࢇཎ≀㉁ࡢᩘࢆቑࡸࡋ࡚ CTA ࢆከࡃᐇ᪋ࡋࠊከࡃࡢࢹ࣮ࢱࢆ཰㞟ࡍࡿࠋࣉࣞࣂࣜࢹ࣮ࢩࣙࣥ◊✲࡛⏝࠸ࡓ⿕㦂≀㉁࡟ࡣ┿ࡢ㠀㑇ఏẘᛶࡀࢇཎ≀㉁ࡢᩘࡀ㑇ఏẘᛶࡀࢇཎ≀㉁࡜ẚ㍑ࡋ࡚ᴟࡵ࡚ᑡ࡞࠿ࡗࡓࡇ࡜࡞࡝ࡀࠊOECD ࡢヨ㦂ἲ࢞࢖ࢻࣛ࢖ࣥ࡜ࡋ࡚ᢎㄆ㸦ྠព㸧ࡉࢀ࡞࠿ࡗࡓ⌮⏤ࡢ୍ࡘ࡜ゝࢃࢀ࡚࠸ࡿࠋࡇࢀࡽࡢ⛉Ꮫⓗ࣭ᢏ⾡ⓗ࡞ၥ㢟Ⅼࡀඞ᭹ࡉࢀࠊࡲࡓṇᘧ࡞ࣂࣜࢹ࣮ࢩࣙࣥ◊✲ࡀᐇ᪋ࡉࢀࢀࡤࠊCTA ࡀࢸࢫࢺ࢞࢖ࢻࣛ࢖ࣥ໬ࡉࢀࡿྍ⬟ᛶࡣ࠶ࡿࠋ
䞉SHE CTA ࡛ࡣࠊᇶᮏⓗ࡞⸆≀௦ㅰᶵ⬟ࢆಖᣢࡋ࡚࠸ࡿ⣽⬊ࢆ⏝࠸ࡿࡇ࡜࠿ࡽࠊ௦ㅰάᛶ໬ࢆᚲせ࡜
ࡍࡿᵝࠎ࡞໬Ꮫ≀㉁ࡢᙧ㉁㌿᥮⬟ࡀ᳨ฟྍ⬟࡛࠶ࡿࠋࡲࡓࠊእ㒊௦ㅰάᛶ໬⣔ࢆ㐺⏝ࡍࡿࡇ࡜࡟ࡼ
ࡾࠊᙧ㉁㌿᥮⬟ࡢቑᙉࡸ᪂ࡓ࡞ᙧ㉁㌿᥮⬟ࢆᣢࡘ໬Ꮫ≀㉁ࢆ᳨ฟ࡛ࡁࡿ࡜ࡋࡓሗ࿌ 18-21) ࡀᑡᩘ࡛ࡣ
࠶ࡿࡀබ⾲ࡉࢀ࡚࠸ࡿࠋࡍ࡞ࢃࡕࠊ㏻ᖖࡢᙧ㉁㌿᥮ヨ㦂࡛㝜ᛶࡢ⤖ᯝࡀᚓࡽࢀࡓ≀㉁࡟㛵ࡋ࡚ࡶࠊእ
㒊௦ㅰάᛶ໬⣔ࢆ㐺⏝ࡍࡿࡇ࡜࡟ࡼࡾࠊ᭦࡟ࡇࡢヨ㦂⣔ࡢṇ☜ᛶࡀ㧗ࡲࡿྍ⬟ᛶࡀ࠶ࡿࠋ

13.

⤖ㄽ

1) CTA ࡣ in vitro ࡟࠾࠸࡚ࠊ໬Ꮫ≀㉁ࢆ᭚㟢ࡍࡿࡇ࡜࡛㉳ࡁࡿࠕ⣽⬊ࡢᙧែࠖࡸࠕ⾲⌧ᙧ㉁ࠖࡢᝏᛶ໬ࢆ
ᣦᶆ࡜ࡋ࡚⭘⒆ㄏⓎᛶࡢ᭷↓ࢆㄪ࡭ࡿヨ㦂ἲࡢ୍ࡘ࡛࠶ࡿࠋ
2) ᙧ㉁㌿᥮ࡋࡓ⣽⬊ࡀᝏᛶ໬ࡍࡿࡇ࡜࡛ࠊࢾ࣮ࢻ࣐࢘ࢫࡸྠ⣔⤫ࡢື≀࡬ࡢ㐀⭘⒆ᛶࡀㄆࡵࡽࢀࡿࡓࡵࠊ
ࡀࢇཎᛶ࡜ࡣఱࡽ࠿ࡢ㛵ಀࡀ࠶ࡿࡇ࡜ࡀ᥎ᐹࡉࢀࡿࠋ
3) ໬Ꮫ≀㉁ࢆ⏝࠸ࡓ CTA ࡢ⤖ᯝ࡜ࠊࡆࡗṑ㢮ࡀࢇཎᛶヨ㦂ࡢ⤖ᯝ࡟ࡣ┦㛵ࡀ࠶ࡿࡇ࡜ࡀሗ࿌ࡉࢀ࡚࠸
ࡿࠋ4) SHE CTA ࡢ⤖ᯝ࡜ࡆࡗṑ㢮࡛ࡢࡀࢇཎᛶヨ㦂࡜ࡢ୍⮴⋡ࠊឤᗘࠊ≉␗ᗘ࡞࡝ࡢᩘ್ࠊ࠾ࡼࡧ IARC ࡢࢢ࣮ࣝࣉูࡢࡀࢇཎ≀㉁࠿ࡽุ᩿ࡍࡿ࡜ࠊSHE CTA ࡣࡀࢇཎᛶヨ㦂ࡢ௦᭰ἲ࡜ࡋ࡚ࡢ఩⨨࡙ࡅ࡛ࡣ࡞ࡃࠊࡀࢇཎ≀㉁᳨ฟࡢࢫࢡ࣮ࣜࢽࣥࢢヨ㦂ἲࡢ୍ࡘ࡜ࡋ࡚఩⨨࡙ࡅࡿࡇ࡜ࡀ࡛ࡁࡿࠋ

23

29

14.

ཧ⪃ᩥ⊩

1)

Guidance Document on the In Vitro Syrian Hamster Embryo (SHE) Cell Transformation Assay, Series on
Testing & Assessment No 214, ENV/JM/MONO (2015)18,OECD (2015), Paris

2)

Report on the Evaluation of Genotoxic or Non-genotoxic Activity of the Organic Chemicals Tested in the SHE
Cell Transformation Assay, 26th Meeting of the Working Group of National Coordinators of the Test Guidelines
Program (WNT), ENV/JM/TG/RD (2014) 17

3)

Pienta, R. J., J. A. Poiley, and W. B. Lebherz III. Morphological transformation of early passage Golden Syrian
embryo cells derived from cryopreserved primary cultures as a reliable in vitro bioassay for identifying divers
carcinogens. Int. J. Cancer: 19, 642-655 (1977)

4)

Watanabe, Masami and Keiji Suzuki, Expression dynamics of transforming phenotypes in X-irradiated Syrian
golden hamster embryo cells. Mutat. Res., 249:71-80 (1991)

5)

Detailed review paper on cell transformation assays for detection of chemical carcinogens, OECD Environment,
Health and Safety Publications, Series on Testing and Assessment, No. 31 OECD (2007)

6)

Report of the Validation Management Team on the ECVAM prevalidation study concerning the SHE pH6.7
Cell Transformation Assay, EC-ECVAM (2010a)

7)

Report of the Validation Management Team on the ECVAM prevalidation study concerning the SHE pH7.0
CTA, EC-ECVAM (2010b)

8)

Results of An Analysis of The Performance of The Syrian Hamster Embryo Cell Transformation Assay at PH
6.7 and PH 7.0, 26th Meeting of the Working Group of National Coordinators of the Test Guidelines Program(WNT), ENV/JM/TG/RD (2014) 16

9)Guidance document on the validation and international acceptance of new or updated test methods for hazard
assessment, OECD Environment, Health and Safety Publications, Series on Testing and Assessment, No. 34
OECD (2005)

10) ESAC Working Group Peer Review Consensus Report on an ECVAM-coordinated prevalidation study
concerning three protocols of the Cell Transformation Assay (CTA) for in vitro carcinogenicity testing (2011)11) EURL ECVAM Recommendation on three Cell Transformation Assays using Syrian Hamster Embryo Cells(SHE) and the BALB/c 3T3 Mouse Fibroblast Cell Line for In Vitro Carcinogenicity Testing (2012)12) Draft Summary Record of a Meeting on Cell Transformation Assay, 14-16th January 2014, Paris, France,
ENV/JM/TG/M(2014)1/REV2

13) Benigni R, Bossa C, Battistelli CL, Tcheremenskaia O, IARC Classes 1 and 2 carcinogens are successfully
identified by analternative strategy that detects DNA-reactivity and celltransformation ability of chemicals.
Mutat. Res., 758 56– 61 (2013)14) Isfort RJ, Kerckaert GA, LeBoeuf RA, Comparison of the standard and reduced pH Syrian Hamster Embryo(SHE) cell in vitro transformation assays in predicting the carcinogenic potential of chemicals. Mutat. Res.,
356: 11-63 (1996)2430

15) LeBoeuf RA, Kerckaert KA, Poiley JA and Raineri R, An interlaboratory comparison of enhanced
morphological transformation of Syrian hamster embryo cells cultured under conditions of reduced bicarbonate
concentration and pH. Mutat. Res., 222: 205-18 (1989)16) Engelhardt G, Schwind KR and Mußler B, The testing of chemicals in Syrian hamster embryo (SHE) celltransformation assay for assessment of carcinogenic potential. Toxicol. in vitro, 18: 213-8 (2004)17) Guidance Document on the In Vitro Bhas42 cells Transformation Assay. Series on Testing & Assessment No
231, ENV/JM/MONO (2016) 1, OECD (2016)18) Tsutsui T, Suzuki N, Maizumi H and Barrett J C, Characterization of an unscheduled DNA synthesis assay with
Syrian hamster embryo cells, Mutat. Res., 129:111-7 (1984)

19) Tsutsui T, Suzuki N, Maizumi H, McLachlan, J A and Barrett J C, Alteration in diethylstilbestrol-induced
mutagenicity and cell transformation by exogenous metabolic activation, Carcinogenesis, 7:1415-8 (1986)20) Tsutsui T, Watanabe E and Barrett J C, Ability of peroxisome proliferators to induce cell transformation,
chromosome aberrations and peroxisome proliferation in cultured Syrian hamster embryo cells, Carcinogenesis,
14: 611-8 (1993)21) Hayashi N, Hasegawa K, Komine A, Tanaka Y, McLachlan J A, Barrett J C and Tsutsui T, Estrogen-induced
cell transformation and DNA adduct formation in cultured Syrian hamster embryo cells, Mol Carcinogenesis,
16:149-56 (1996)25

31