triple labeling staining for breast cancer tissue final

8/6/2019 Triple Labeling Staining for Breast Cancer Tissue Final

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Identifying breast cancer stem cell in theprimary tumour

Presented By:

Jifei Zhao


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Br east cance r

B reast cancer is the most common type of cancer among female allover the world, it affect nearly 10% of women.

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Normal Breast Tissue T he arrowpoints to a cross-section of a duct(1).

And Lobules (2)

Normal Breast Tissue --T he arrow points toa cross-section of a duct. Lobules can beseen off to the left


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T he TNM classification is based on the size of the tumor ( T ), whether or not the tumor has spread to the lymph nodes ( N ) in the armpits, andwhether the tumor has metastasized ( M )

Stage 0 is a pre-cancerous or marker condition, either ductal carcinomain situd (DCIS) or lobular carcinoma in situ (LCIS).

Stages 13 are defined as 'early' cancer with a good prognosis.

Stage 4 is defined as 'advanced' and/or 'metastatic' cancer with a poor prognosis.

TNM classification


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W hat is DCIS

D uctal hyperplasia means that too many cells are present

Atypical ductal hyperplasia means that there are too manycells (hyperplasia) and they are starting to take on an abnormalappearance

D uctal carcinoma in situ means that there are too many cellsand they have the features of cancer, but stay inside of the duct.

DCIS-MI (DCIS with microinvasion) means that a fewof the cancer cells have started to break through the wallof the duct. T his is considered to be a slightly moreserious form of DCIS.

Invasive ductal cancer means that the cancer cells havebroken beyond the breast duct. T he breast cancer is nolonger a DCIS but an invasive ductal carcinoma, the mostcommon type of breast cancer.


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T riple staining for breast cancer tissue

CD44+/CD24- is commonly used as the cell surface marker for cancer stem cells.

B ut there is a need for more accurate target to the specific stemlike cells.

CK19 is another breast cancer marker and imply to metastasis.So we apply a triple labeling for CD44 CD24 CK19


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CD44 and its variant isforms

Presence of CD44 variant e on in different N eoplastic cell lines

CD44v6 and breast cancer:

Shiptisin(2009) performedthe I C staining for breasttissue using CD44V6antibody, and found that

most of the stromal cells arenegative for CD44V6.

Cd44v6 in normal tissue(left) and in IDC(right)


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Compare CD44 and CD44V6

Representative immunohistochemical findings of normal breast tissue ( N ) and invasive ductalcarcinoma of the breast (C) using M abs anti-CD44 , anti-CD44v3, and anti-CD44v6. Infiltratinglymphocytes, which stain with anti-CD44 but are nonreactive to anti-CD44v3 and anti-CD44v6,are used as internal controls (arrows).


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secondary Antibody PrimaryAntibody

Ex(nm) Em(nm) Catalog #

Alex Fluor 594 goatanti-mouse SFR(H+L)

Ck19-MousemonoclonalIgG2a(light

chain)

590 617 A31631

Alex Fluor488 goat anti rabbitSFX kit (H+L)

Cd24- rabbitpolyclonal IgG(abgentap8782a)

495 519 A31619

Streptavidin, Alexa

Fluor 546 conjugate

CD44/ CD44v6Biotin-labeledab

(ebioscienceBMS125BT)

556 573 A11056

T he antibody table


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Spectra view of three dyes

Spectra viewer of ale a fluor 488, 546,594


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Reference

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5, Shipitsin M , Campbell LL, Argani P, et al. M olecular definition of breast tumor heterogeneity. Cancer Cell (2007)11:259273.

6, Xavier Roca, et al, CD44 Isoform E pression Follows T wo Alternative Splicing Pathways in B reast T issue. American Journal of Pathology/(1998) 153:183-190


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triple labeling staining for breast cancer tissue final

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