Time dependent
16. doxorubicin کberberineלՁဍዦޑלဍዦࢲ܄
! ! ᇘႵࢂΕ 1x107SCC-4 ಒझࡕǴҺཀϩԋΟಔ(ಔϤଫ)Ǵಃ
ಔࢂڋಔޑᇘႵ๏ϒݙ 30ȝlDMSOǴಃΒಔޑᇘႵ๏ϒα ᗯ१doxorubicin (4mg/kg)ǴಃΟಔޑᇘႵ๏ϒαᗯ१ berberine (10 mg/kg)Ǵவ໒ۈډಃ 10 ϺဍዦΕࡕکុԿჴᡍֹԋ(34 Ϻ)Ǵဍ
ዦӧЁޑᡏᑈЁκکελ (ߏ৩کอ৩)ࢂຼᒵΒԛǴෳໆ ဍዦޑߏ৩Ȑaȑکอ৩ȐbȑǴਥᏵϦԄ V=1/2ab2 ीᆉဍዦᡏᑈǶ
ዦ=(ჹྣಔဍዦᡏᑈɡݯᕍಔဍዦᡏᑈ)ʏჹྣಔဍዦᡏᑈ
×100%Ƕीᆉ่݀ᡉҢ doxorubicin ޑဍዦޑᡏᑈܴᡉޑКڋಔૈ
ᇨว 65%ޑဍዦڋǴberberine ޑဍዦޑᡏᑈܴᡉޑКڋಔૈᇨ ว52%ޑဍዦڋ(ӵკ 4-16)Ƕ
(A)
კ 4-16 ౦ᅿҜ౽ނኳࠠޑဍዦᡏᑈڋ
ಃϖകʳ ፕ
ಃҽ ೱનёၸcaspase-8, -9ک-3 җAIFᆶEndoGၡ৩ᇨ วΓᜪՁᕎಒझਲ਼SCC-4ಒझΫޑϩηբҔᐒᙯϐࣴز
! ! ᕎੱࢂΓᜪЬाޑԝΫচӢǴځٰ่݀Ծܭಒझߞ৲ሀӦ
ՉϩፓಒझޑቚکӸࢲᆶᡂ౦ 91Ǵᇨวᕎੱಒझౢғಒझ
ΫࢂᕎੱݯᕍޑЬाౣǶԿܭځϩΕಒझޑѦӧၡ৩܈җಈ ጕᡏࢂคݤޕၰޑ92ǵ93ǵ94ǶಒझΫၡ৩хࡴ caspase-8, -9, -3 ک -12 کಈጕᡏၡ৩ࢂੋډಒझՅન C ޑࢬрᆶٰԾܭಈጕᡏډझஏᾕ
ԋ Apaf-1 ک caspase-9 ޑಒझΫᡏԖᜢ95ǶჴሞΒచၡ৩Ξϕ
࣬רԶፄᚇᜤᒣǴԖ٤ൔςჴނϯᏢނ፦ӧჹלόӕޑᕎ
ੱБय़ڀԖወӧޑϯᏢႣٛᆶϯᏢᕍݤޑբҔǶೱનᡉҢӧ٤ᜪ
ࠠޑᕎಒझԖᇨวಒझࢥ܄ޑբҔǴӢԜ،ۓҔೱનჹΓᜪՁᕎಒ झ SCC-4 ಒझࢂցૈᇨวಒझΫࢂҁჴᡍޑࣴزҞޑǶ
! ! ҁჴᡍ่݀ᡉҢೱનૈ෧ϿಒझޑӸࢲǴёૈҗܭಒझΫ
ԋಒझԝΫǶჴೱનᇨวΓᜪՁᕎ SCC-4 ಒझਲ਼ޑಒझΫǴ
ၗჴૈቚу sub-G1 DNA ޑኧໆǴDAPI ᑻӀࢉՅΨᡉҢೱન ᇨวΓᜪՁᕎಒझޑ DNA ্Ƕ
! ! ࣴزೱનԋԖᜢಒझΫޑϩηᐒᙯޑԖᜢচӢǴಒ झΫ࣬ᜢೈқޑ߄ၲǴፏӵǽBcl-2ǵ Bcl-xL ǵ Bax ǵBad ǵ Bakǵ
caspase-3ǵ-8 ک-9ǵcytochrome cǵApaf-1ǵFasǵFADD کಒझጢႝ
ՏӧΓᜪՁᕎ SCC-4 ಒझਲ਼ຑǶBax/Bcl-2 ޑ෧ϿКޑёૈচ ӢࢂಒझጢႝՏޑ഼ѨǴ่݀ԋ cytochrome c ޑញܫᏤठಒझΫ
96Ƕ่݀ࢂவࢬԄಒझሺޑϩჴೱનૈ෧ϿಒझጢႝՏǴՋБ ᗺᏀݤޑ่݀ᡉҢೱનૈගଯ cytochrome c ޑНѳǴගϲಒझΫ
ೈқǴፏӵǽBcl-2 ک Bcl-xLǶԖኧᏵࡰр Bax ૈϩಈጕᡏጢ ޑֹکಈጕᡏၸ܄೯ϾޑԋǴ่݀วғ cytochrome c ޑញ ܫ97ǶӃޑπբᡉҢಈጕᡏၸ܄೯ϾՔᒿಈጕᡏଏཱུϯǶ ڥ֎ڋ܈ڈᐟǴ୷፦εǴ୷፦ஏᾕਡੳለᅰکጢϣೈқញܫǴ хࡴ cytochrome c98Ƕ
! ! ಒझԝΫௗڙၡ৩ૈૈࢲϯ/ှ caspase-8 ਸӦፓǴࢂᗖ
่ಒझԝΫௗӝηޑ܄کಒझԝΫௗڙՔᒿࢲϯ caspase-8 ک caspase-3 วғಒझΫ 99Ƕࣴز่݀ࡰрೱનӧΓᜪՁᕎ SCC-4 ಒझԋ caspase- 8, -9 ک -3 ှቚуǴ٠Ъቚу caspase-9 ှᏤ ठᒿࡕӧΠෞޑ caspase-3 ࢲϯǴ(ঁಒझΫޑՉޣ)Ƕ่݀ᡉ Ңೱનૈߦ Apaf-1 ޑ߄ၲǴᏤठߦ caspase-3 ࢲϯЇଆಒझ
ΫǶ
! ! ࣴز،ۓࢂٗঁሀੋϷೱનᇨวಒझΫǴҔ caspase ᏊೀǴฅࡕಒझᎦӧԖೱનӸӧύǴ่݀ࡰр
pan-caspase inhibitorࢂঁߔЗcaspaseೱનޑᇨวԖࡐமޑբ ҔǴฅࡕڋcaspase-3, caspase-8, ک caspase-9Ƕ೭٤่݀ᡉҢ
ೱનೀޑΓᜪՁᕎSCC-4 ಒझǴᇨวಒझΫޑࢲϯሡा೭٤ caspaseǴѬࢂFasޑख़ाڋᏊ(לFasޑSCC-4ಒझೀ) ೱનೀ
ޑಒझܴᡉޑ෧ϿಒझΫǴᡉҢೱનᇨวಒझΫӧࢌᅿำࡋ
ಒझԝΫௗڙၡ৩ፓǶ
ಃΒҽ ೱનёၸ FAK, IKK, NF-țB, u-PA ک MMP-2 ک -9 ၡ৩ڋΓᜪՁᕎ SCC-4 ಒझਲ਼ޑᙯ౽ᆶߟΕբҔᐒᙯϐࣴز
! ! ೱનޑלᕎբҔςӧӭόӕᜪࠠޑᕎੱкϩჴ 50ǵ51ǵ54ǵ
55ǵ57-60
Ǵ่݀ჴೱનૈ෧ϿΓᜪՁᕎ SCC-4 ಒझޑӸࢲǴςӧ Ӄޑࣴز၁ॊၸǶฅԶೱનӧΓᜪՁᕎ SCC-4 ಒझޑᙯ౽کߟ Εޑࢲ܄کߞ৲ၡ৩ᗋ҂ԖࣴزൔǴวೱનӧΓᜪՁᕎ SCC-4 ಒझૈεεӦ෧ե ERK 1/ 2 ޑᕗለکࢲϯǴERK 1/ 2 ᇡࣁჹ NF-țB ޑᨿϯߚதख़ा 100ǴNF-țB ჹܭፓဍዦಒझޑቚǵಒझΫک ठᕎ୷Ӣߚதख़ा 101ǴӭӃޑࣴزᡉҢೱનޑלဍዦࢲ܄ё
ૈԖϩᆶၸڋ NF-țB ޑࢲϯޑፓԖᜢ 102-105Ǵ٠Ъೱન ӧΓᜪՁᕎ SCC-4 ಒझᗨฅڋ NF-țB ՠΨᇨว IțBǴӢԜ่݀
ᡉҢೱનޑלᕎࢲ܄ёૈᆶ ERK1/2 MAPK ک NF-țB ޑߞ৲ၡ৩ ԖᜢǶ
! ! ғᏢکੰಔᙃख़༟ၸำǴٯӵǺαᘰӝǵखजΕೌǵဍ ዦߟΕǵᙯ౽کՈᆅཥғࢂᆶ୷፦ߎឦೈқ䁙 (MMPs) ࣬ᜢǴᄬԋ
ঁϩݜৎǶᎋ٩ᒘ܄ϣᴏ䁙ࢂሡा ECM ޑफ़ှբҔ 106-108Ƕ MMP-9 ࢂ NF-țB ޑΠෞႢǴૈफ़ှ ECMǴׯᡂಒझ-ಒझکಒ झ ECM ໔ޑቹៜǴکᕎಒझޑߟΕԖᜢ 109ǶECM ڋᏊςჴ
ૈߔЗϣҜಒझޑࢲϯǴჹཥՈᆅޑቚکߟΕ110ǵ111ࢂόё܈લޑ
ाનǴЇଆ൳ঁ MMPs ޑ୷ጢϩှǴӢԜ MMP-2 ک MMP-9 ՟Яჹ
୷ጢޑಃ IV ࠠጤচޑफ़ှߚதख़ा 112-114ǴMMP-2 ޑ߄ၲၟဍዦߟ ΕǵՈᆅཥғǵᙯ౽کӆғ࣬ᜢ 115ǵ116Ƕ
! ! MMP-2 ک MMP-9 ૈᒣלᕎᛰނޑݯᕍႢǴҗܭӧܴጤΒ ᅿ䁙ޑफ़ှբҔࢂ୷ጢၨεޑԋҽǴMMP-2 ک MMP-9 ޑೈқ፦
ڋߟҍǵߟΕ܈ဍዦᙯ౽ޑ߄ 117-119ǶᡉҢೱનૈ෧ϿΓᜪՁᕎ SCC-4 ಒझޑ MMP-2 ک MMP-9 ޑೈқ፦ໆǴԖൔࡰр MMP-2 ک MMP-9 ޑೈқ፦ک uPA ሡा NF-țB ک AP-1120ǵ121Ǵೱનӧ NF-țB
ೈқ፦ޑڋբҔૈᇥܴڋ MMP-2 ک MMP-9 ޑ୷Ӣ߄ၲ 122ǵ
123Ƕ܌ڬޕ ERK ࢲϯஒڈᐟΒঁԄբҔፓϡҹǴхࡴ AP-1
ک NF-țB ޑᗖ่Տ֟Ǵӧፓ MMP-9 ޑ୷Ӣ߄ၲՔᄽঁख़ाޑف ՅǶ
! ! ӧҞޑൔೱનૈ෧Ͽ NF-țB ޑೈқ፦ໆǴՠΨගଯΑ IțB
ޑೈқ፦ໆǴёૈᏤठڋ MMP-2 ک MMP-9 ޑೈқ፦ໆǶೱન
ૈڀᡏӦڋ NF-țB ޑࢲϯբҔǴ IțB (NF-țB ޑڋԛൂϡ) ޑफ़ှբҔߔЗǴЪૈߔЗ NF-țB ӛಒझਡ่ᄬޑᙯᒵ 124ǴԖൔ
൳ঁלᙯ౽ፄӝނڋ MMPǴڋբҔᆶ൳ঁόӕޑᐒᙯ࣬ᜢ
125Ǵ೭٤ᐒᙯޔௗڋ MMP ޑࢲϯ䁙Ǵߔᛖ MMP ޑࢲϯک෧Ͽ MMPޑ୷Ӣ߄ᄽϷೈқ፦ໆǶ
ಃΟҽ ೱનჹΓᜪՁᕎ SCC-4 ಒझਲ਼ӧᇘႵࢲᡏϣޑϩηբ Ҕᐒᙯϐࣴز
! ! ᗨฅӭᏵςᡉҢೱનӧӭΓᜪᕎಒझਲ਼ᇨวಒझຼ
යଶᅉکᇨวಒझΫǴ٠ЪӧࢲᡏϣޑނኳࠠתᄽלᕎբҔ52ǵ57ǵ
126-129
ǶฅԶᗋ҂ԖൔᡉҢೱનӧΓᜪՁᕎᡒރಒझӧࢲᡏϣޑ౦ ᡏҜ౽ႵޑቹៜǶҁჴᡍӧҞޑࣴزࢂಃԛගٮӧࢲᡏϣೱ નჹΓᜪՁᕎᡒރಒझ SCC-4 ᕎڀԖфਏޑᏵǶ SCC-4 ಒझς
٬ҔӭԃǴჹܭࣴزՁᕎᡒރಒझӵӕঁኳࠠಒझਲ਼129ǵ130Ƕ
! ! ҁჴᡍύࢲᡏϣޑࣴزวೱનӧ౦ᡏҜ౽ኳࠠ 10 mg/kg
ૈڋဍዦޑғߏǴ೭٤ᆶٰԾܭӧࢲᡏϣޑೱન ( 30 ȝM ) ޑ
ঁ࣬՟ᐚࡋޑࣴزԖั༾ӦόӕǴӧSCC-4 ಒझޑಒझቚԖᡉ
ޑಒझࢥનբҔ ( Їଆຬၸ 50% ޑಒझԝΫ ) کᇨวಒझ
ΫǶό۩ӦǴӧҞޑࣴزؒԖीᆉೱનӧဍዦಔᙃޑжᖴ
ᐚࡋǴฅԶဍዦӧԴႵൂᐱௗڙೱનࢂКڋಔεऊϿ52%Ǵӧ
Չೱન๏ᛰؒԖᡉӦࢥ܄բҔǴܴӧᡏख़کѦᢀಞ܄٠ؒԖׯ ᡂǶ
! ! аԜБԄޑೱનӧ SCC-4 ࢲᡏϣޑ౦ᡏҜ౽ғߏڋբ Ҕࢂϸࢀӧࢲᡏϣ܌ᛘளޑ่݀Ǵ࣬ჹӦǴೱન10 mg/kg ೀޑ ᇘႵК 4mg/kg doxorubicinΞКڋಔᕇள׳λޑᡏᑈȐӵკ4-15ȑǶ εऊ65% ޑڋǶဍዦௗڙೱનೀុғߏࢂᄌޑǴࡰр SCC-4 ಒ झ ޑ ౦ ᡏ Ҝ ౽ ޑ ֹ ӄ ଏ ϯ ค ݤ Ҕ ൂ ޑ ೀ ၲ ԋ ( berberine ܈ doxorubicin )Ƕ೭ΨࡰрٗᅿፄӝԄޑೀჹၲډֹӄ ޑϸᔈёૈࢂሡाޑǶќѦޑӢનΨஒԵቾǴٯӵǺp53 ǴԿϞ p53
ࢂന೯தޑँᡂဍዦޑڋ୷ӢǴکલЮᐒૈޑ p53 ᆶቚуဍዦғ ߏޑӒᓀ࣬ᜢǴηᓍᕎ!Ca Ski cell ࢂঁ p53 ޑ positive cells Ƕ Ѭࢂൔޤဏᕎ H1299 cellsޑ౦ᡏҜ౽ޑဍғߏКӧԴႵޑޤ ᕎA549 cells کೱનӧϯᏢᕍݤޑфਏp53-positive-A549 tumor xenograft К p53-deficient-H1299 tumor xenograft ࢂ׳ܴᡉޑ131Ƕ
! ! ӧҁࣴزჴᡍύࢲᡏϣᆶࢲᡏѦΒޣϐ໔ޑৡ౦Ǵځ่݀ёૈᆶ ӧࢲᡏϣКӧࢲᡏѦӸӧόӕޑжᖴԄԖᜢǴೱનک/܈жᖴނ ᆶಒझғߏΨԖڋբҔǴ೭ёૈϸࢀӧೱન෧ϿSCC-4ဍዦғ ߏޑᛰΚک/܈ᛰਏϕ࣬ቹៜ࣬ᜢǶ
ಃϤക ่ፕ
! ! ҁፕЎ२ӃፕೱનӧࢲᡏѦޑՁᕎಒझϩηሀၡ৩ᇨว ಒझΫޑբҔᐒᙯǴځԛೱનӧࢲᡏѦޑՁᕎಒझޑᙯ౽ᡂ ϯᆶԋ౽ᆶߟΕޑϩηբҔᐒᙯǴനࡕаೱનӧᇘႵࢲ ᡏϣჹՁᕎಒझޑڋբҔᐒᙯǶҁፕЎޑ่ፕϷวӵΠǺ
1. ೱનёၸcaspase-8, -9ک-3 җAIFᆶEndoGၡ৩ᇨวΓᜪ ՁᕎSCC-4ಒझਲ਼ಒझΫޑϩηբҔᐒᙯϐࣴز
! ! ೱનࢂᅿᾳނϯᏢނ፦ޑಔԋǴӧΓᜪՁᕎಒझ SCC-4 ᇨ ᇨวಒझΫǶFas (ௗӝೈқ) Քᄽೱનߦ௴ۈ caspase-8 ک caspase-9 ԾှکࢲϯޑঁௗڙᏔǴ௴ۈ caspase ှکࢲϯբ Ҕว caspase-3Ƕcaspase-3 ᇨวݩޑӭኬ܄ǴനಖᏤठಒझ
Ϋ ( ӵკ 6-1 )Ƕ
კ 6-1 ೱનჹ SCC-4 ಒझਲ਼ᇨวಒझΫၡ৩Ƕ
2. ೱનёၸ FAK, IKK, NF-țB, u-PA ک MMP-2 ک -9 ၡ৩
ڋΓᜪՁᕎ SCC-4 ಒझਲ਼ޑᙯ౽ᆶߟΕբҔᐒᙯϐࣴز
! ! ೭ࢂԖࣴزኧᏵಃԛࡰрೱનڋΓᜪՁᕎSCC-4 ಒझਲ਼ ޑಒझᙯ౽کߟΕࢂၸ p-ERK ک NF-țBޑߞ৲ၡ৩ᏤठMMP-2 ک-9ޑڋ ( ӵკ6-2 )Ƕ٠ЪᡉҢೱનڋ FAK, p-38, p-JNK ک p-ERK ᏤठAP-1 کNF-țB วғբҔǴڋMMP-2 ک-9.ޑ߄
ၲǶೱનᔈёԵቾբࣁڋՁᕎޑᙯ౽کߟΕϐݯᕍᛰނǴ҂ٰޑ
ࣴزஒሡाຑೱનݯᕍᕎੱޑወӧ܄Ƕ
კ 6-2 ೱનڋ SCC-4 ಒझਲ਼ޑಒझᙯ౽کߟΕၡ৩Ƕ
3. ೱનჹΓᜪՁᕎಒझਲ਼ SCC-4 ӧᇘႵࢲᡏϣޑϩηբҔᐒᙯϐ
ࣴز
! ! ೱન 10 mg/kg ჴࡼҗѤϺԛޑဎ๚ݙǴӧՁᕎ౦ᡏҜ
౽ԴႵኳࠠޑՁSCC-4ဍዦ෧ϿғߏࢂԖਏޑǴ่݀ᡉҢ berberine ޑဍዦޑᡏᑈܴᡉޑКڋಔૈᇨว 52%ޑဍዦڋǴ೭ঁว߄ ҢೱનޑಃԛޑࣴزჴᡍࢂԖਏޑǴӧঁՁᕎԴႵ౦ᡏҜ౽
ኳࠠύೱન൩ӵӕঁՁᕎޑႣٛᛰނǶ
!
კ 6-3 ೱનڋᇘႵ SCC-4 ဍዦޑڋǶ Ȑ*** pɦ0.001ǴڀԖीᡉНྗȑ
ୖԵЎ
1. Չࡹଣፁғ.୯ΓΜεԝӢ.҇୯ΐΜϖԃΜΒДΜѤВϦ
2. ۟ǺဍዦᖏੱഢाǴޕॣрހޗǴѠч 2003ǹp.1-146 3. ៝դநǺύᙴѦࣽᏢǴޕॣрހޗǴѠч 1989ǹp.205 4. మǷଯޚ໋ǺᅬࣽЈளǴ॥рހޗǴѠч 1976ǹp.15
5. మǷֆᖰጓǺᙴےߎ᠘ǴཥЎᙦрހϦљǴѠч 1985ǹ3Ǻp.210
6. మǷЀ४ǺЀМോࣽઝਜǴ؇ߎ㔬ᙴᏢӄਜǴύ୯ύᙴᛰрހޗǴ ч٧ 1999ǹp.458
7. మǷх҉ੀǺკຏോࣽࡰඓǴཥЎᙦрހǴѠч 1976ǹ3Ǻp.7 8. మǷ៝ШዂǺᅬᙴεӄǴ॥рހޗǴѠч 1973ǹ15Ǻp.4 9. ॕϘӸǺύᙴဍዦᏢǴࣽᏢрހޗǴч٧ 1983ǹp.224-5
10. ӀൺਜֽጓᒠǺύՋᙴ่ӝဍዦᏢǴӀൺਜֽǴѠч 1992ǹ p.335 11. ۟ǺဍዦੰǴΓ҇ፁғрހޗǴч٧ 1982ǹp.45
12. ݅ࡿ۪ǺύՋᙴੰӜჹྣεᜏڂǴΓ҇ፁғрހޗǴч٧ 2002ǹ p.429-30
13. ഋ᎒ుǺ.жύᙴဍዦᏢǴΓ҇ፁғрހޗǴч٧ 2003ǹp.306 14. ΟߍЎϯǺύᛰჴҔკڂǴΟߍрހϦљǴѠч 2008ǹp.278-9.
15. ؼҡጓǺҁᆜҞқ၉ᆒǴϣᆾђࣽמрހޗǴهঢ় 2004ǹ p.113.
16. మǷበᒠǺઓၭҁǴύᙴђᝤрހޗǴч٧ 1987ǹp.69.
17. ఉǷഏѶඳǺӜᙴձᒵǴΓ҇ፁғрހޗǴч٧ 1986ǹp.116 18. ֺǷႨǺҁკǴӼᔇࣽמрހޗǴч٧ 1994ǹp.125 19. ֺǷஊےበǺҁ़ကǴহЎၗрހޗǴѠύ 1987ǹڔ 8 p.2 20. ϡǷЦӳђǺ෯నҁǴহЎၗрހޗǴѠύ 1987ǹp.98 21. ܴǷਔࣔǺҁᆜҞǴШЎϯрހޗǴѠࠄ 2000ǹp.447.
22. ܴǷϟᇯǺඳۢӄਜǴѠᖄ୯॥рހޗǴѠч 1980ǹp.917 23. మǷ؋ܳǺҁഢाǴЎӀკਜрހޗǴѠч 1984ǹp.178 24. మǷ⻡ǺҁচǴύ୯ύᙴᛰрހޗǴч٧ 1996ǹp.44 25. మǷذǺҁǴֻਜֽǴѠч 1974ǹp.190
26. మǷယϺγǺҁှǴϖࢪрހޗǴѠч 1984ǹp.54 27. మǷഋঅ༜Ǻઓၭҁ᠐ǴדᇻਜֽǴѠч 1987ǹp.31 28. మǷ࠴ǺҁϩǴੇࣽמрހޗǴੇ 1989ǹp.71
29. ஷўጓǺҁᆜҞқ၉ᆒҁǴεӦрހޗǴѠч 2008ǹp.142.
30. Parkin DM, Pisani P, Ferlay J. Estimates of the worldwide incidence of 25 major cancers in 1990. Int J Cancer 1999; 80: 827-41.
31. Hashibe M, Brennan P, Benhamou S, Castellsague X, Chen C, Curado MP, L. Dal Maso, Daudt AW,Fabianova E, Fernandez L, Wunsch-Filho V, Franceschi S, Hayes RB, Herrero R, Koifman S, La Vecchia C, Lazarus P, Levi F, Mates D, Matos E, Menezes A, Muscat J, Eluf-Neto J, Olshan AF, Rudnai P, Schwartz SM, Smith E, Sturgis EM, Szeszenia-Dabrowska N, Talamini R, Wei Q, Winn DM, Zaridze D, Zatonski W, Zhang ZF, Berthiller J, Boffetta P. Alcohol drinking in never users of tobacco, cigarette smoking in never drinkers, and the risk of head and neck cancer:
pooled analysis in the International Head and Neck Cancer Epidemiology Consortium, J Natl Cancer Inst 2007; 99: 777-89.
32. Rahman M, Sakamoto J, Fukui T. Calculation of population attributable risk for bidi smoking and oral cancer in south Asia, Prev Med 2005; 40: 510-4.
33. Balaram P, Sridhar H, Rajkumar T, Vaccarella S, Herrero R, Nandakumar A, Ravichandran K, Ramdas K, Sankaranarayanan R, Gajalakshmi V, Munoz N, Franceschi S. Oral cancer in southern India:
the influence of smoking, drinking, paan-chewing and oral hygiene, Int J Cancer 2002; 98: 440-5.
34. Room R, Babor T, Rehm J. Alcohol and public health, Lancet 2005;
365: 510-30.
35. Garidou A, Tzonou A, Lipworth L, Siqnorello LB, Kalapothaki V, Trichopoulos D. Life-style factors and medical conditions in relation to esophageal cancer by histologic type in a low-risk population. Int J Cancer 1996; 68: 295-9.
36. Franceschi S, Bidoli E, Baron AE, Barra S, Talamini R, Serraino D, Lavecchia C. Nutrition and cancer of the oral cavity and pharynx in north-east Italy. Int J Cancer 1991; 47: 20-5.
37. Lam AK. Molecular biology of esophageal squamous cell carcinoma.
Crit Rev Oncol Hematol 2000; 33: 71-90.
38. Baan R, Straif K, Grosse Y, Secretan B, El Ghissassi F, Bouvard V, Altieri A, Cogliano V. Carcinogenicity of alcoholic beverages, The lancet oncology 2007; 8: 292-3.
39. Parkin DM, Bray F. Chapter 2: The burden of HPV-related cancers, Vaccine 2006; 3: S11-25.
40. Pintos J, Franco EL, Kowalski LP, Oliveira BV, Curado MP. Use of
wood stoves and risk of cancers of the upper aero-digestive tract: a case-control study. Int J Epidemiol 1998; 27: 936-40.
41. Velly AM, Franco EL, Schlecht N, Pintos J, Kowalaki LP, Oliveira BV, Curado MP. Relationship between dental factors and risk of upper aerodigestive tract cancer. Oral Oncol 1998; 34: 284-91.
42. Applebaum KM, Furniss CS, Zeka A, Posner MR, Smith JF, Bryan J, Eisen EA, Peters ES, McClean MD, Kelsey KT. Lack of association of alcohol and tobacco with HPV16-associated head and neck cancer, J Natl Cancer Inst 2007; 99: 1801-10.
43. Boccia S, Cadoni G, Sayed-Tabatabaei FA, Volante M, Arzani D, De Lauretis A, Cattel C, Almadori G,. van Duijn CM, Paludetti G, Ricciardi G.
CYP1A1, CYP2E1, GSTM1, GSTT1, EPHX1 exons 3 and 4, and NAT2 polymorphisms, smoking, consumption of alcohol and fruit and vegetables and risk of head and neck cancer, J cancer Res clin oncol 2008;
134: 93-100.
44. Franco EL, Kowalski LP, Oliveira BV, Curado MP, Pereira R, Silva ME, Fava AS.Torloni H. Risk factors for oral cancer in Brazil: a case-control study. Int J Cancer 1989; 43: 992-1000.
45. Schlecht NF, Franco EL, Pintos J, Negassa A, Kowalski LP, Oliveira BV, Curado MP. Interaction between tobacco and alcohol consumption and the risk of cancers of the upper aero-digestive tract in Brazil. Am J Epidemiol 1999; 150: 1129-37.
46. Hong WK, Sporn MB. Recent advances in chemoprevention of cancer.
Science 1997; 278: 1073–7.
47. Kucuk O. Chemoprevention of prostate cancer. Cancer Metastasis Rev 2002; 21: 111–24.
48. Craig WJ. Phytochemicals: guardians of our health. J Am Diet Assoc 1997; 97: S199–204.
49. Schiller LR. Review article: anti-diarrhoeal pharmacology and therapeutics. Aliment Pharmacol Ther 1995; 9: 87-106.
50. Hwang JM, Kuo HC, Tseng TH, Liu JY, Chu CY. Berberine induces apoptosis through a mitochondria/caspases pathway in human hepatoma cells. Arch Toxicol 2006; 80: 62-73.
51. Iizuka N, Miyamoto K, Okita K, Tangoku A, Hayashi H, Yosino S, Abe T, Morioka T, Hazama S, Oka M. Inhibitory effect of Coptidis
Rhizoma and berberine on the proliferation of human esophageal cancer cell lines. Cancer Lett 2000; 148: 19-25.
52. Nishino H, Kitagawa K, Fujiki H, Iwashima A. Berberine sulfate inhibits tumor-promoting activity of teleocidin in two-stage carcinogenesis on mouse skin. Oncology 1986; 43: 131-4.
53. Sack RB, Froehlich JL. Berberine inhibits intestinal secretory response of Vibrio cholerae andEscher ichia coli enterotoxins. Infect Immun 1982;
35: 471–5.
54. Fukuda K, Hibiya Y, Mutoh M, Koshiji M, Akao S, Fujiwara H.
Inhibition of activator protein 1 activity by berberine in human hepatoma cells. Planta Med 1999; 65: 381-3.
55. Fukuda K, Hibiya Y, Mutoh M, Koshiji M, Akao S, Fujiwara H.
Inhibition by berberine of cyclooxygenase-2 transcriptional activity in human colon cancer cells. J Ethnopharmacol 1999; 66: 227-33.
56. Lin JG, Chung JG, Wu LT, Chen GW,Chang H, Wang TF. Effects of berberine on arylamine N-acetyltransferase activity in human colon tumor cells. Am J Chin Med 1999; 27: 265-75.
57. Peng PL, Hsieh YS, Wang CJ, Hsu JL, Chou FP. Inhibitory effect of berberine on the invasion of human lung cancer cells via decreased productions of urokinase-plasminogen activator and matrix metallo- proteinase-2. Toxicol Appl Pharmacol 2006; 214: 8-15.
58. Kim SA, Kwon Y, Kim JH, Muller MT,Chung IK. Induction of topoisomerase II-mediated DNA cleavage by a protoberberine alkaloid, berberrubine. Biochemistry 1998; 37: 16316-24.
59. Letasiova S, Jantova S, Cipak L, Muckova M. Berberine-anti proliferative activity in vitro and induction of apoptosis/necrosis of the U937 and B16 cells. Cancer Lett 2006; 239: 254-62.
60. Mantena SK, Sharma SD, Katiyar SK. Berberine inhibits growth, induces G1 arrest and apoptosis in human epidermoid carcinoma A431 cells by regulating Cdki-Cdk-cyclin cascade, disruption of mitochondrial membrane potential and cleavage of caspase 3 and PARP. Carcinogenesis 2006; 27: 2018-27.
61. Mantena SK, Sharma SD, Katiyar SK. Berberine, a natural product, induces G1-phase cell cycle arrest and caspase-3-dependent apoptosis in human prostate carcinoma cells. Mol Cancer Ther 2006; 5: 296-308.
62. Lin JG, Chung JG, Wu LT, Chen GW, Chang HL, Wang TF. Effects of berberine on arylamine N-acetyltransferase activity in human colon tumor cells, The Am J Chin Med 1999; 27: 265-75.
63. Fukuda K, Hibiya Y, Mutoh M, Koshiji M, Akao S, Fujiwara H.
Inhibition by berberine of cyclooxygenase-2 transcriptional activity in human colon cancer cells, J Ethnopharmacol 1999; 66: 227-33.
64. Ho YT, Lu CC, Yang JS, Chiang JH, Li TC, Lin JG, Chung JG.
Berberine induced apoptosis via promoting the expression of caspase-8, -9 and -3, apoptosis inducing factor and endonuclease G in SCC-4 human tongue squamous carcinoma cancer cells, Anticancer Res. Submitted.
65. Ho YT, Yang JS, Li TC, Lin JJ ,Lin JG, Lai KC, Ma CY, W. Wood WG , Chung JG, Berberine suppresses in vitro migration and invasion of human SCC-4 tongue squamous cancer cells through the inhibitions of FAK, IKK, NF-țB, u-PA and MMP-2 and -9, Cancer Lett. 2009; 279:
155-62.
66. HoYT, Yang JS, Lu CC, Chiang JH, Li TC, Lin JJ, Lai KC, Liao CL, Lu HF, Lin JG, ChungJG, Berberine Inhibits Human Tongue Squamous Carcinoma Cancer Tumor Growth in a Murine Xenograft Model , Phytomedicine. 2009; Epub ahead of print.
67. Cohen MH, Gootenberg J, Keegan P, Pazdur R. FDA drug approval summary: bevacizumab (Avastin) plus Carboplatin and Paclitaxel as first-line treatment of advanced/metastatic recurrent nonsquamous non-small cell lung cancer.Oncologist 2007; 12: 713-8.
68. [No authors listed] Drug combination approved for advanced breast cancer.FDA Consum 2002; 36: 5.
69. Wu SN, Yu HS, Jan CR, Li HF,Yu CL. Inhibit ry effects of berberine on voltage 2 and calcium 2 activated potassium currents in human myeloma cells. Life Sci 1998; 62: 2283-94.
70. Kettmann V, Kosfalova D, Jantova S, Cernakova M, Drimal J. In vitro cytotoxicity of berberine against HeLa and L1210 cancer cell lines.
Pharmazie 2004; 59: 548-51.
71. Mitani N, Murakami K, Yamaura T, Ikeda T, Saiki I. Inhibitory effect of berberine on the mediastinal lymph node metastasis produced by orthotropic implantation of Lewis lung carcinoma. Cancer Lett 2001; 165:
35-42.
72. Kuo CL, Chi CW, Liu TY. The anti-inflammatory potential of berberine in vitro and in vivo. Cancer Lett 2004; 203: 127-37.
73. Chi CW , Chang YF, Chao TW ,Chiang SH, P’eng Fk, Lui WY, Liu TY. Flowcytometric analysis of the effect of berberine on the expression of glucocorticoid receptors in human hepatoma HepG2 cells Cell. Life Sci 1994; 54: 2099-107.
74. Kuo CL , Chou CC, Yung BY. Berberine comp lexes w ith DNA in the berberine 2 induced apoptosis in human leukemic HL- 260 cells. Cancer Lett 1995; 93: 1992-200.
75. Iizuka N ,Miyamoto K, Okita K, Tangoku A, Havashi H,Yosino S,Abe T,Morioka T, Hazama S. Inhibitory effect of Coptidis Rhizoma and berberine on the proliferation of human esophageal cancer cell lines.
Cancer Lett 2000; 148: 19-25.
76. Mitani N ,Murakam i K, Yamaura T, Ikeda T, Saiki I. Inhibito ry effect of berberine on the mediastinal lymphnode metastasis produced by orthotopic implantation of Lewis lung carcinoma. Cancer Lett 2001; 165:
45-52.
77. Anis KV, Rajeshkumar NV, Kuttan R. Inhibition of chemical carcinogensis by berberine in rats and mice. J Pharm Pharmacol 2001; 53:
763-8.
78. Hartwell LH, Kastan MB. Cell cycle control and cancer. Science 1994;
266: 1821-8.
79. Nigg EA. Cyclin-dependent protein kinases: key regulators of the eukaryotic cell cycle. Science 1995; 17: 471-80.
80. Hunter T, Pines J. Cyclins and cancer. II: Cyclin D and CDK inhibitors come of age. Cell 1994; 79: 573-82.
81. Ohtsubo M, Theodoras AM, Schumacher J,Robert JM,Pagono M.
Human cyclin E, a nuclear protein essential for the G1-to-S phase transition. Mol Cell Biol 1995; 15: 2612-24.
82. Girard F, Strausfeld U, Fernandez A, Lamb N. Cyclin A is required for the onset of DNA replication in mammalian fibroblasts. Cell 1991; 67:
1169-79.
83. Chen J, Takson PK, Kirschner MW, Dutta A. Separate domains of p21 involved in the inhibition of Cdk kinase and PCNA. Nature 1995; 374:
84. Fuller GM, Shields D, Molecular basis of medical cell biology, McGraw-Hill publishing, 1998; 1: 113.
85. Raff MC, Barres BA, Burne JF, Coles HS, Ishizaki Y, Jacobson MD.
Programmed cell death and the control of cell survival: lessons from the nervous system. Science 1993; 262: 695-700.
86. Behl C. Apoptosis and Alzheimer's disease. Nature Transm 2000; 107:
1325-44.
87. Fadok VA, Voelker DR, Campbell PA, Cohen JJ, Bratton DL, Henson PM. Exposure of phosphatidylserine on the surface of apoptotic lymphocytes triggers specific recognition and removal by macrophages.
Immunol 1992; 148: 2207-16.
88. Hanahan D, Weinberg RA. The hallmarks of cancer. Cell 2000; 100:
57-70.
89. Poon RT, NqIO, Lau C, Zhu LX, Yu WC, Lo CM, Fan ST, Wong J.
Serum vascular endothelial growth factor predicts venous invasion in hepatocellular carcinoma: a prospective study. Ann Surg 2001; 233:
227-35.
90. Varmus H, Weinberg RA, Genes and the biology of cancer, W.H.
Freeman&Co 1992; 1: 180.
91. Ashkenazi A, Dixit VM. Death receptors: signaling and modulation.
Science 1998; 281: 1305-8.
92. Mehmet H. Caspases find a new place to hide. Nature 2000; 403:
29-30.
93. Shi Y. Mechanisms of caspase activation and inhibition during apoptosis. Mol Cell 2002; 9: 459-70.
94. Yang J, Liu X, Bhalla K, Kim CN, Ibrado AM, Cai J, Peng TI, Jones DP, Wang X. Prevention of apoptosis by Bcl-2: release of cytochrome c from mitochondria blocked. Science 1997; 275: 1129-32.
95. Bouchier-Hayes L, Lartigue L, Newmeyer DD. Mitochondria:
pharmacological manipulation of cell death. J Clin Invest 2005; 115:
2640-7.
96. Bernardi P, Krauskopf A, Basso E, Petronilli V, Blachlv-Dvson E, Di Lisa F, Forte MA. The mitochondrial permeability transition from in vitro artifact to disease target. FEBS J 2006; 273: 2077-99.
97. Hengartner MO. The biochemistry of apoptosis. Nature 2000; 407:
770-6.
98. Liu X, Yue P, Zhou Z, Khuri FR, Sun SY. Death receptor regulation and celecoxib-induced apoptosis in human lung cancer cells. J Natl Cancer Inst 2004; 96: 1769-80.
99. Mantena SK, Sharma SD, Katiyar SK. Berberine, a natural product, induces G1-phase cell cycle arrest and caspase-3-dependent apoptosis in human prostate carcinoma cells, Mol cancer ther 2006; 5: 296-308.
100. Nakano H, Shindo M, Sakon S, Nishinaka S, Mihara M, Yagita H, Okumura K. Differential regulation of IkappaB kinase alpha and beta by two upstream kinases, NF-kappaB-inducing kinase and mitogen-activated protein kinase/ERK kinase kinase-1, Pro Natl Acad Sci USA 1998; 95:
3537-42.
101. Orlowski RZ, Baldwin AS Jr. NF-kappaB as a therapeutic target in cancer, Trends Mol Med 2002; 8: 385-9.
102. Pandey MK, Sung B, Kunnumakkara AB, Sethi G, Chaturvedi MM, Aggarwal BB. Berberine modifies cysteine 179 of IkappaBalpha kinase, suppresses nuclear factor-kappaB-regulated antiapoptotic gene products, and potentiates apoptosis, Cancer Res 2008; 68: 5370-9.
103. Yi P, Lu FE, Xu LJ, Chen G, Dong H, Wang KF, Berberine reverses free-fatty-acid-induced insulin resistance in 3T3-L1 adipocytes through targeting IKKbeta, World J Gastroenterol 2008; 14: 876-83.
104. Hu JP, Nishishita K, Sakai E, Yoshida H, Kato Y, Tsukuba T, Okamoto K. Berberine inhibits RANKL-induced osteoclast formation and survival through suppressing the NF-kappaB and Akt pathways, Eur J Pharmacol 2008; 580: 70-9.
105. Enk R, Ehehalt R, Graham JE, Bierhaus A, Remppis A, Greten HJ.
Differential effect of Rhizoma coptidis and its main alkaloid compound berberine on TNF-alpha induced NFkappaB translocation in human keratinocytes, J Ethnopharmacol 2007; 109: 170-5.
106. Woessner JF Jr, Gunja-Smith Z. Role of metalloproteinases in human osteoarthritis, J Rheumatol Suppl 1991; 27: 99-101.
107. Ray JM, Stetler-Stevenson WG. The role of matrix metalloproteases and their inhibitors in tumour invasion, metastasis and angiogenesis, Eur Respir J 1994; 7: 2062-72.
108. Stetler-Stevenson WG. Matrix metalloproteinases in angiogenesis: a moving target for therapeutic intervention, J Clin Inves 1999; 103:
1237-41.
109. Stallings-Mann M, Radisky D. Matrix metalloproteinase-induced malignancy in mammary epithelial cells, Cells Tissues Organs 2007; 185:
104-10.
110. Murphy AN, Unsworth EJ, Stetler-Stevenson WG. Tissue inhibitor of metalloproteinases-2 inhibits bFGF-induced human microvascular endothelial cell proliferation, J Cell Physiol 1993; 157: 351-8.
111. Benelli R, Adatia R, Ensoli B, Stetler-Stevenson WG, Santi L, Albini A. Inhibition of AIDS-Kaposi's sarcoma cell induced endothelial cell invasion by TIMP-2 and a synthetic peptide from the metalloproteinase propeptide: implications for an anti-angiogenic therapy, Oncol Rese 1994;
6: 251-7.
112. Stetler-Stevenson WG. Type IV collagenases in tumor invasion and metastasis, Cancer Metastasis Rev 1990; 9: 289-303.
113. Giancotti FG, Ruoslahti E. Integrin signaling, Science 1999; 285:
1028-32.
114. Zeng ZS, Cohen AM, Guillem JG. Loss of basement membrane type IV collagen is associated with increased expression of metalloproteinases 2 and 9 (MMP-2 and MMP-9) during human colorectal tumorigenesis, Carcinogenesis 1999; 20: 749-55.
115. Wang M, Wang T, Liu S, Yoshida D, Teramoto A. The expression of matrix metalloproteinase-2 and -9 in human gliomas of different pathological grades, Brain Tumor Pathol 2003; 20: 65-72.
116. Komatsu K, Nakanishi Y, Nemoto N, Hori T, Sawada T, Kobayashi M.
Expression and quantitative analysis of matrix metalloproteinase-2 and -9 in human gliomas, Brain Tumor Pathol 2004; 21: 105-12.
117. Bianco FJ Jr, Gervasi DC, Tiguert R, Grignon DJ, Pontes JE, Crissman JD, Fridman R, Wood DP Jr. Matrix metalloproteinase-9 expression in bladder washes from bladder cancer patients predicts pathological stage and grade, Clin Cancer Res 1998; 4: 3011-6.
118. Cockett MI, Murphy G, Birch ML, O'Connell JP, Crabbe T, Millican AT, Hart IR, Docherty AJ, Matrix metalloproteinases and metastatic cancer, Bioche Soc symp 1998; 63: 295-313.
119. Papathoma AS, Zoumpourlis V, Balmain A, Pintzas A. Role of matrix metalloproteinase-9 in progression of mouse skin carcinogenesis, Mol Carcinog 2001; 31: 74-82.
120. Bond M, Fabunmi RP, Baker AH, Newby AC, Synergistic upregulation of metalloproteinase-9 by growth factors and inflammatory cytokines: an absolute requirement for transcription factor NF-kappa B, FEBS Lett 1998; 435: 29-34.
121. Farina AR, Tacconelli A, Vacca A, Maroder M, Gulino A, Mackay AR. Transcriptional up-regulation of matrix metalloproteinase-9 expression during spontaneous epithelial to neuroblast phenotype conversion by SK-N-SH neuroblastoma cells, involved in enhanced invasivity, depends upon GT-box and nuclear factor kappaB elements, Cell Growth Differ 1990; 10: 353-67.
122. Lin TH, Kuo HC, Chou FP, F. Lu J. Berberine enhances inhibition of glioma tumor cell migration and invasiveness mediated by arsenic trioxide, BMC Cancer 2008; 8: 58.
123. Wartenberg M, Budde P, De Marees M, Grunheck F, Tsang SY, Huang Y, Chen, ZY, Hescheler J, Sauer H, Inhibition of tumor-induced angiogenesis and matrix-metalloproteinase expression in confrontation cultures of embryoid bodies and tumor spheroids by plant ingredients used in traditional chinese medicine, Lab Invest 2003; 83: 87-98.
124. Shishodia S, Potdar P, Gairola CG, Aggarwal BB. Curcumin (diferuloylmethane) down-regulates cigarette smoke-induced NF-kappaB activation through inhibition of IkappaBalpha kinase in human lung epithelial cells: correlation with suppression of COX-2, MMP-9 and cyclin D1, Carcinogenesis 2003; 24: 1269-79.
125. Hidalgo M, Eckhardt SG. Development of matrix metalloproteinase inhibitors in cancer therapy, J Natl Cancer Inst 2001; 93: 178-93.
126. Yount G, Qian Y, Moore D, Basila D, West J, aldape K, Arrold N,Shalev N, Haas-Kogan D. Berberine sensitizes human glioma cells, but not normal glial cells, to ionizing radiation in vitro . J Exp Ther Oncol 2004; 4: 137–43.
127. Lin HL, Liu TY, Wu CW, Chi CW. Berberine modulates expression of mdr1 gene product and the responses of digestive track cancer cells to Paclitaxel. Br J Cancer 1999; 81: 416–22.
128. Choi BH, Ahn IS, Kim YH, Park JW,Lee SY,Hvnn CK,Do MS.
Berberine reduces the expression of adipogenic enzymes and inflammatory molecules of 3T3–1 adipocyte. Exp Mol Med 2006; 38:
599–60
129. Yang SF, Yang WE, Chang HR, Chu SC, Hsieh YS. Luteolin induces apoptosis in oral squamous cancer cells. J Dent Res. 2008; 87: 401-6.
130. Lin YT, Yang JS, Lin HJ, Tan TW, Tang NY, Chaing JH, Chang YH, Lu HF, Chung JG. Baicalein induces apoptosis in SCC-4 human tongue cancer cells via a Ca2+-dependent mitochondrial pathway. In vivo, 2007;
21: 1053-8
131. Katiyar SK, Meeran SM, Katiyar N, Akhtar S. p53 cooperates berberine-induced growth inhibition and apoptosis of non-small cell human lung cancer cells in vitro and tumor xenograft growth in vivo. Mol Carcinog. 2009; 48: 24-37.
Molecular mechanisms of growth inhibition on human tongue cancer SCC-4 cells by berberineǺin vitro and in vivo
Yung-Tsuan Ho
Major professor ȅTsai-Chung Li
Graduate Institute of Chinese Medical Science, China Medical University 1. Berberine induced apoptosis via promoting the expression of caspase-8, -9 and -3, apoptosis inducing factor and endonuclease G in SCC-4 human tongue squamous carcinoma cancer cells
Phytochemicals have been recognized to have potential chemo-preventive or chemotherapeutic efficacy in cancer treatment. Here, we hypothesized that berberine would have anti-cancer activities in human tongue cancer SCC-4 cells. Results indicated that berberine decreased the viability of SCC-4 cells which was initiated by the generation of reactive oxygen species, via an increase in cytosolic Ca2+. Berberine induced apoptosis was associated with a reduction of the mitochondrial membrane potential associated with changes in the Bax/Bcl-2 ratio, release of cytochrome c from mitochondria and activation of down-stream caspase-3. Real time PCR showed that berberine stimulated gene expression of caspase-8, -9 and -3, apoptosis-inducing factor (AIF) and Endo G. The present study demonstrated that berberine-mediated apoptosis of SCC-4 cells is
regulated by ROS, mitochondria, caspase-3 -dependent and -independent pathways, suggesting that berberine may be considered for future studies as a promising therapeutic candidate for human tongue cancer.
2. Berberine suppresses in vitro migration and invasion of human