Identification and applications of MGAgs

-------a group of novel gastric cancer associated antigens

Institute of Digestive Diseases ,Xijing Hospital, Fourth Military Medical University, Xi’an, China

Daiming Fan,Xueyong Zhang,Xitao Chen, ,Jialu Hu, Zhengxian Mu , Jun Ren, Jie Ding,Kaichun Wu

Gastric cancer

• Cause about 200,000 deaths annually

• Early detection is not very easy

• Mass survey is not feasible in China

• Cause about 200,000 deaths annually

• Early detection is not very easy

• Mass survey is not feasible in China

Positive immunohistochemical reaction (% of cases)Positive immunohistochemical reaction (% of cases)

The nature and immunohistochemical reactivity of MG series monoclonal

antibodies

The nature and immunohistochemical reactivity of MG series monoclonal

antibodies

MGa-5MGa-7MGa-9MGa-11MGb-1MGb-2MGc-1MGd-1MGe-1MGe-2MGg-1MGg-2

MGa-5MGa-7MGa-9MGa-11MGb-1MGb-2MGc-1MGd-1MGe-1MGe-2MGg-1MGg-2

IgG1IgG1IgG2IgG1IgG1IgG1IgG1IgG1IgG1IgMIgG1IgG1

IgG1IgG1IgG2IgG1IgG1IgG1IgG1IgG1IgG1IgMIgG1IgG1

28.666.772.636.068.842.050.070.020.046.744.867.6

28.666.772.636.068.842.050.070.020.046.744.867.6

16.725.025.0 016.7 0 9.1 0 040.920.532.3

16.725.025.0 016.7 0 9.1 0 040.920.532.3

25.022.230.016.7 014.3 0 0 055.6 013.0

25.022.230.016.7 014.3 0 0 055.6 013.0

000000000000

000000000000

000000000000

000000000000

mAbmAb Igsubclass

Igsubclass Gastric

cancerGastriccancer

Coloniccancer

Coloniccancer

Esophagealcancer

Esophagealcancer

Hepaticcancer

Hepaticcancer

Pancreaticcancer

Pancreaticcancer

Early detection of gastric cancer from precancerous lesions of stomach

Early detection of gastric cancer from precancerous lesions of stomach

DysplasiaDysplasia

RetrogressRetrogress

Malignant transformationMalignant transformation

DisappearDisappear

4.0~19.0%4.0~19.0%

Follow up study of patients with dysplasia of gastric mucosa

Follow up study of patients with dysplasia of gastric mucosa

Dysplasia(90 cases)Dysplasia(90 cases)

Stained by MGa-7Stained by MGa-7

29 cases (+)29 cases (+)

Malignant change in 6 cases( 4 cases

early gastric cancer)

Malignant change in 6 cases( 4 cases

early gastric cancer)

16 cases (42 months)16 cases (42 months)

61 cases (-)61 cases (-)

No malignant change but 1 case became positive in staining

No malignant change but 1 case became positive in staining

19 cases (41 months)

19 cases (41 months)

Immunofluorescent diagnosis of cancer cells in malignant ascitic fluid or

pleural effusion

Immunofluorescent diagnosis of cancer cells in malignant ascitic fluid or

pleural effusionMethodsMethods

Immunoflourescent examinationConventional cytological examination

Immunoflourescent examinationConventional cytological examination

CasesCases

76

76

76

76

Positivecases

Positivecases

69

59

69

59

%%

90.7

77.6

90.7

77.6

(P<0.05)(P<0.05)

Radioimmunoimaging of tumor in vivo

• The satisfactory imaging results were obtained

in 73.3% of patients with gastric cancer(74/10

1). Metastatic tumor in liver, pancreas and sup

raclavicular lymph node could also be detected.

• The satisfactory imaging results were obtained

in 73.3% of patients with gastric cancer(74/10

1). Metastatic tumor in liver, pancreas and sup

raclavicular lymph node could also be detected.

RII of Patients with Gastric CancerRII of Patients with Gastric Cancer

131I-McAb

MG7F(ab)2

MG7IgG

MGd1

MGb2

MG11

MG7+MGd1

131I-McAb

MG7F(ab)2

MG7IgG

MGd1

MGb2

MG11

MG7+MGd1

Positive rate(%)

87.5(7/8)

75.9(22/29)

44.4(4/9)

50.0(2/4)

0 (0/7)

100(3/3)

Positive rate(%)

87.5(7/8)

75.9(22/29)

44.4(4/9)

50.0(2/4)

0 (0/7)

100(3/3)

Detection of MGAgs for serological diagnosis of gastric cancer by

immuno-PCR

Detection of MGAgs for serological diagnosis of gastric cancer by

immuno-PCR

0%

20%

40%

60%

80%

100%

0%

20%

40%

60%

80%

100%

EIAEIAEIAEIA RIARIARIARIA IRMAIRMAIRMAIRMA

Detection rate of gastric cancer using existing serological diagnostic methodsDetection rate of gastric cancer using

existing serological diagnostic methods

GC antigensGC antigensGC antigensGC antigens

Gastric lumenGastric lumenGastric lumenGastric lumen Blood circulationBlood circulationBlood circulationBlood circulation

metabolizedmetabolized metabolizedmetabolized

Reasons of low dectection rate for gastric cancer

Reasons of low dectection rate for gastric cancer

95%95%95%95% 5%5%5%5%

????

IFIFIFIF EIAEIAEIAEIA RIARIARIARIA

• High sensitivityHigh sensitivity

• 25,000 publications25,000 publications

• Just for detection of nucleic acidJust for detection of nucleic acid

• High sensitivityHigh sensitivity

• 25,000 publications25,000 publications

• Just for detection of nucleic acidJust for detection of nucleic acid

Advantages and disadvantages of PCR

Advantages and disadvantages of PCR

Immuno-PCR

Immuno-PCR

pXJ 19 pXJ 19MG7 -p XJ 19

KATO III MGKATO III MG77AgAg PP

(x cell/ml) concentration(x cell/ml) concentration

IPCRIPCR 2 x 10 2 x 101 1 3.8 x 103.8 x 10 －－ 14 14 <0.001<0.001

EIA EIA 2 x 102 x 105 5 3.0 x 103.0 x 10 －－ 11 11

KATO III MGKATO III MG77AgAg PP

(x cell/ml) concentration(x cell/ml) concentration

IPCRIPCR 2 x 10 2 x 101 1 3.8 x 103.8 x 10 －－ 14 14 <0.001<0.001

EIA EIA 2 x 102 x 105 5 3.0 x 103.0 x 10 －－ 11 11

Sensitivity comparison of IPCR to EIA

Sensitivity comparison of IPCR to EIA

Detection of MGAgs in different gastric diseases with IPCR

Cases Positive Cases Positive ％％

Gastric cancer 198 164 Gastric cancer 198 164 82.882.8

Peptic ulcer 78 6 7.7Peptic ulcer 78 6 7.7

Chronic gastritis 118 Chronic gastritis 118 7 5.9 7 5.9

Donor 236 Donor 236 2 0.8 2 0.8

Positive results of MGAgs in different cancers with IPCR

Cancer of Cases Positive Cancer of Cases Positive ％ ％ Stomach 198 164 Stomach 198 164 82.882.8Colon 90 Colon 90 40 44.4 40 44.4Esophagus Esophagus 86 15 17.4 86 15 17.4Ovary Ovary 45 45 1 2.2 1 2.2Lung Lung 66 4 6.1 66 4 6.1 Liver Liver 84 0 0 84 0 0Uterus Uterus 27 0 27 0 00

Positive results of MGAgs in different histological patterns of gastric cancer

Histological typeHistological type

Well differentiatedWell differentiated

Poorly differentiatedPoorly differentiated

Mucous adenocarcinomaMucous adenocarcinoma

Mucous cell carcinomaMucous cell carcinoma

Cases Positive %Cases Positive %

151 122 80.8 151 122 80.8

80 56 70.080 56 70.0

31 20 64.531 20 64.5

29 16 29 16 55.2 55.2

Positive results of MGAgs at different stages of gastric cancers

Stage Cases Positive %Stage Cases Positive %

Early 31 12 38.7Early 31 12 38.7

Advanced 116 91 78.4Advanced 116 91 78.4

Comparison of MGAgs to other tumor markers

MarkerMarker

CEA CEA

CA50 CA50

CA19-9 CA19-9

TAG-72 TAG-72

MGMG77 Ag-IPCR Ag-IPCR

％％

12.0 12.0

7.27.2

7.27.2

8.48.4

8.4 8.4

Benign diseases Benign diseases (83)(83)

10 10

6 6

6 6

7 7

7 7

％ ％

33.7 33.7

27.9 27.9

38.4 38.4

45.3 45.3

81.481.4

Gastric ca. Gastric ca.

(86) (86)

29 29

24 24

33 33

39 39

70 70

10u

100u

1000u1u

10 万种药

MDR

抗癌药

多个已知耐药相关分子在胃癌中的表达

MDR1 10%

MRP 12%

LRP 10%

GST-pi 36%

Non-detected

42%

Total ratios 58%

42% 58%

Sensitive cells

MDR cells

polyA+ RNAs proteins

SSH/DD-PCR 2-D gel electrophoresis

83 genes 30 proteins

Anticancer drug

In vitro selection

SGC7901 SGC7901/VCR

pH 10 - 3 IEF

SD

S --P

AG

E

pH 10 - 3 IEF

Protein spectrums displayed by 2-D gel

SGC7901 SGC7901/VCR

11 11

19 19

12 1211

19

30 proteins

3 specifically expressed in MDR cells

6 up-regulated in MDR cells

19 down-regulated in MDR cells

2 specifically expressed in sensitive cells

新耐药分子 MGr1-Ag 的发现

Accession number

AF503367

MGr1

Fourteen molecules have been proved to play certain roles in tumor MDR:

8. mitotic arrest deficient 2

9. Occludin

10.phosphotriose isomerase

11.Ribosomal protein L6

12.putative magnesium transporter

13.novel gene PTD001

14.novel gene V62

1. MGr1-Ag

2. prion protein

3. Ribosomal protein S13

4. Ribosomal protein L23

5. zinc ribbon protein ZNRD1

6. calcyclin binding protein

7. tumor susceptibility gene 101

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