HPV DNA RNA 蛋白表达与宫颈癌发病的人群研究

HPV

DNA

,

m

RNA和E6蛋白表达与宫颈癌发病的人群研究内

容HPV

DNA、mRNA和E6蛋白表达的相互关系不同宫颈病变程度人群中HPV DNA、mRNA和E6蛋白表达的差异不同HPV

DNA、mRNA和E6蛋白表达人群CIN2+的患病风险背

景子宫颈癌自然史人乳头瘤病毒（HPV）长控区（LCR）:不具有编码功能早期区：病毒的复制和转录E1，E2，E4，E5，E6，E7晚期区：衣壳蛋白的编码L1，L2DoorbarJ.Thebiologyandlife-cycleofhumanpapillomaviruses[J].Vaccine,2012,30

Suppl5:F55-F70.HPV

与宫颈病变的关系JD.Thepapillomaviruslifecycle.[J].JournalofClinicalVirologytheOfficialPublicationofthePanAmericanSocietyforClinicalVirology,2005,32suppl

1(3):7–15.不同情况下HPV

相关分子表达的情况DNA/mRNA/蛋白宫颈正常CIN1CIN2CIN3/宫颈癌基因维持增殖期基因扩增装配释放E1+/+/++/+/+++/++/++++/-/-++/+/+++-/+-/+--/-/-E2+/+/++/+/+++/++/++++/-/-++/+/+++-/+-/+--/-/-E4+/-/-+/-/-++/++/++++/++/++++/+/+++-/+-/+-+-/-/-E5+/-/-+/-/-++/++/++++/-/-++/+/+++-/+-/+-+-/-/-E6+/-/-+/++/++++/+/+++/-/-++/++/+++++/+++/+++++++/++++/++++E7+/-/-+/++/++++/+/+++/-/-++/++/+++++/+++/+++++++/++++/++++L1+/-/-+/-/-++/-/-++/++/++++/-/-++-/+-/+-++-/-/-L2+/-/-+/-/-++/-/-++/++/++++/-/-++-/+-/+-+-/-/-JD.Thepapillomaviruslifecycle.[J].JournalofClinicalVirologytheOfficialPublicationofthePanAmericanSocietyforClinicalVirology,2005,32suppl

1(3):7–15.研究对象分组研究现场合计宫颈癌CIN2/3CIN1/良性病变正常中国医学科学院肿瘤医院501050700810山西省肿瘤医院150302080280四川大学华西第二附属医院150302080280天津市中心妇产科医院150302080280958065110350合计59518017510502000检测方法HPV

DNA：cobas4800

HPV检测系统

(Roche)，检测14种HR-HPV，并对HPV16和HPV18进行分型A. 扩增曲线的cut-off

?B.

标准曲线:

ct值和log10最初拷贝数作图E6/E7

mRNA：Aptima

HPV

mRNA检测技术

(Hologic)，检测14种HR-HPV的E6/E7mRNA表达情况3份重复的阳性校准品（CO）3份阴性校准品（NEG）单孔标本（S），读数为：S/CO；外加质控（IC）APTIMA

检测结果 标准阴性分析物S/CO

<

0.50IC≥IC临界值IC≤2,000,000

RLU阳性分析物S/CO

≥

0.50IC≤2,000,000

RLU分析物≤

13,000,000

RLUE6癌蛋白：OncoE6检测技术

(Arbor

Vita

Corporation)，检测HPV16或HPV18的E6蛋白表达情况研究进展HPV

DNATotalNegativeHPV16(+) HPV18(+)HR-HPV(+) HR-HPV(+)mRNATotalNegativePositiveN35730327E6癌蛋白TotalNegativeHPV16

(+)HPV18(+)N1432113227082HPV

DNA，mRNA和蛋白表达的相互关系结果HPV

DNA与mRNA检测信号强度的关系不同型别HPV

DNA拷贝数与mRNA表达的关系HPV16

DNA与E6癌蛋白表达的关系HPV16DNA

LoadNegative Low HighTotalHPV16E6

(-)1025

(98.0%)106

(55.2%)30

(15.5%)1161

(81.1%)HPV16E6

(+)21

(2.0%)86

(44.8%)163

(84.5%)270

(18.9%)Total1046

(100.0%)192

(100.0%)193

(100.0%)1431

(100.0%)HPV18

DNA与E6癌蛋白表达的关系HPV18DNA

LoadNegative Low HighTotalHPV18E6

(-)1316

(96.3%)20(62.5%)13

(39.4%)1349

(94.3%)HPV18E6

(+)50

(3.7%)12

(37.5%)20

(60.6%)82

(5.7%)Total1366

(100.0%)32

(100.0%)33

(100.0%)1431

(100.0%)HPV

DNA拷贝数与E6癌蛋白信号强度的关系HPV

DNA、mRNA和E6癌蛋白表达的相互关系HPVDNA

(-)HPVDNA

(+)mRNA(-)mRNA(+)TotalmRNA(-)mRNA(+)TotalHPV16

E6(-)20

(90.9%)3

(60.0%)23

(85.2%)7

(87.5%)100

(31.1%)107

(32.4%)HPV16

E6(+)2

(9.1%)2

(40.0%)4

(14.8%)1

(12.5%)222

(68.9%)223

(67.6%)HPV18

E6(-)22

(100.0%)3

(60.0)25

(92.6%)8

(100.0%)247

(76.7%)255

(77.3%)HPV18

E6(+)0

(0.0%)2

(40.0%)2

(7.4%)0

(0.0%)75

(23.3%)75

(22.7%)Total22

(100%)5

(100%)27

(100%)8

(100%)322

(100%)330

(100%)HPV16,

18

DNA、mRNA和E6癌蛋白表达的相互关系HPV16DNA

(-)HPV16DNA

(+)mRNA(-)mRNA(+)TotalmRNA(-)mRNA(+)TotalHPV16

E6(-)23

(92.0%)63

(78.8%)86

(81.9%)4

(80.0%)40

(16.2%)44

(17.5%)HPV16

E6(+)2

(8.0%)17

(21.3%)19

(18.1%)1

(20.0%)207

(83.8%)208

(82.5%)Total25

(100.0%)80

(100.0%)105

(100.0%)5

(100.0%)247

(100.0%)252

(100.0%)HPV18DNA

(-)HPV18DNA

(+)mRNA(-)mRNA(+)TotalmRNA(-)mRNA(+)TotalHPV18

E6(-)29

(100.0%)244

(83.6%)273

(85.0%)1

(100.0%)6

(17.1%)7

(19.4%)HPV18

E6(+)0

(0.0%)48

(16.4%)48

(15.0%)0

(0.0%)29

(82.9%)29

(80.6%)Total19

(100.0%)292

(100.0%)321

(100.0%)1

(100.0%)35

(100.0%)36

(100.0%)结

论HPV

mRNA的表达量与HPV

DNA含量正相关HPV

E6癌蛋白的表达量随着HPV

DNA含量增加呈上升趋势无论何种HPV

DNA表达状况，mRNA阳性人群中E6癌蛋白的阳性率均高于mRNA阴性人群不同宫颈病变程度人群中HPV DNA、mRNA和蛋白表达的差异纳入人群的病理诊断NormalCIN1CIN2CIN3CancerTotalN(%) 721

(59.6%)60

(5.0%)43

(3.6%)66

(5.5%)319

(26.4)1209

(100%)不同宫颈病变程度人群中HPV

DNA拷贝数不同宫颈病变程度人群中mRNA阳性率CIN1CIN2CIN3CancerTotalP

trendmRNA

(-) 4

(66.7%)3

(33.3%)1

(6.7%)19

(6.0%)27

(7.8%)mRNA

(+) 2

(33.3%)6

(66.7%)14

(93.3%)299

(94.0%)321

(92.2%)<0.001Total6

(100.0%)9

(100.0%)15

(100.0%)318

(100.0%)348

(100.0%)不同宫颈病变程度人群中E6癌蛋白阳性率NormalCIN1CIN2CIN3CancerTotalHPV16

E6(-)714

(99.0%)55

(91.7%)37

(86.0%)36

(54.5%)HPV16

E6(+)7

(1.0%)5

(8.3%)6

(14.0%)30

(45.5%)106(33.2%) 948

(78.4%)P

trend<0.001213(66.8%) 261

(21.6%)HPV18

E6(-)718

(99.6%)58

(96.7%)43

(100.0%)64

(97.0%)HPV18

E6(+)3

(0.4%)2

(3.3%)0

(0.0%)2

(3.0%)246(77.1%) 1129

(93.4%)P

trend<0.00173

(22.9%) 80

(6.6%)HPV16/18

E6(-)712

(98.8%)54

(90.0%)37

(86.0%)35

(53.0%)HPV16/18

E6(+)9

(1.2%)6

(10.0%)6

(14.0%)31

(47.0%)81

(25.4%) 919

(76.0%)P

trend<0.001238(74.6%) 290

(24.0%)Total721

(100.0%)60

(100.0%)43

(100.0%)66

(100.0%)319(100.0%) 1209

(100.0%)不同宫颈病变程度人群中E6癌蛋白的信号强度NormalCIN1CIN2CIN3CancerTotalNormal714

(99.0%)55

(91.7%)37

(86.0%)36

(54.5%)106

(33.2%)948

(78.4%)12

(0.3%)2

(3.3%)4

(9.3%)10

(15.2%)38

(11.9%)56

(4.6%)HPV16234

(0.6%)1

(0.1%)1

(1.7%)1

(1.7%)0

(0.0%)2

(4.7%)8

(12.1%)4

(6.1%)44

(13.8%)43

(13.5%)57

(4.7%)51

(4.2%)40

(0.0%)1

(1.7%)0

(0.0%)4

(6.1%)29

(9.1%)34

(2.8%)50

(0.0%)0

(0.0%)0

(0.0%)4

(6.1%)59

(18.5%)63

(5.2%)Normal718

(99.6%)58

(96.7%)43

(100.0%)64

(97.0%)246

(77.1%)1129

(93.4%)12

(0.3%)0

(0.0%)0

(0.0%)1

(1.5%)38

(11.9%)41

(3.4%)HPV18230

(0.0%)1

(0.1%)1

(1.7%)1

(1.7%)0

(0.0%)0

(0.0%)1

(1.5%)0

(0.0%)11

(3.4%)9

(2.8%)13

(1.1%)11

(0.9%)40

(0.0%)0

(0.0%)0

(0.0%)0

(0.0%)3

(0.9%)3

(0.2%)50

(0.0%)0

(0.0%)0

(0.0%)0

(0.0%)12

(3.8%)12

(1.0%)Total721

(100.0%)60

(100.0%)43

(100.0%)66

(100.0%)319

(100.0%)1209

(100.0%)不同宫颈病变程度人群中HPV DNA、mRNA和HPV16/18

E6癌蛋白表达的差异DNA/mRNA/E6Total+/+/++/+/-+/-/++/-/--/+/+-/+/--/-/+-/-/-CIN10

(0.0%)2

(33.3%)0

(0.0%)0

(0.0%)0

(0.0%)0

(0.0%)0

(0.0%)4

(66.7%)6

(100.0%)CIN22

(22.2%)2

(22.2%)0

(0.0%)0

(0.0%)0

(0.0%)2

(22.2%)0

(0.0%)3

(33.3%)9

(100.0%)CIN38

(53.3%)5

(33.3%)0

(0.0%)1

(6.7%)0

(0.0%)1

(6.7%)0

(0.0%)0

(0.0%)15

(100.0%)Cancer233

(73.3%)61

(19.2%)1

(0.3%)3

(0