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Breast Cancer Associated
Fibroblast(BCAFs) :
Microenviroment.
CHUNGNAM NATIONAL UNIVERSITY
MEDICAL SCHOOL of Biochemistry
PJL Lab.
Hong Bum Park
Breast cancer and Fibroblast cell line by co-culture on media
Tumourmicro-environment elicits innate resistance
to RAF inhibitors through HGF secretion
Nature Ref#-0001
Clinical and functional significance of loss of caveolin-1
expression in breast cancer-associated fibroblasts
J. Pathology Ref#-0002
DMEM
High Glucose
Sodium Pyruvate
GlutaMAX™
Phenol Red
MEMa
Ribonucleosides
Deoxyribonucleosides
Phenol Red
L-glutamine
Minimum Essential Medium (MEM) that contains non-essential amino acids, sodium pyruvate, lipoic acid,
vitamin B12, biotin, and ascorbic acid
- We show that cancer associated fibroblast co-culture be used DMEM media.
-ATCC suggest breast fibroblast cell line was usually EMEM, so we are choice media by DMEM, but primary
breast cancer cell was not.
- DMEM, MEM and RPMI are different in composition. when we are choose media, we are usually search of
on ATCC. If we select wrong media, cells can be change in proliferation, migration ability and morphology.
Breast cancer cell lines and Stromal cell lines
Tumourmicro-environment elicits innate resistance
to RAF inhibitors through HGF secretion
Nature Ref#-001
Breast cancer cell line
BT-474
EFM192A
HCC1419
HCC1569
MCF-7
Breast
Breast
Breast
Breast
Breast
ATCC
DSMZ
ATCC
ATCC
ATCC
HTB-20
ACC 258
CRL-2326
CRL-2330
HTB-22
MDA-MB-175-VII
MDA-MB-361
SK-BR-3
ZR-75-1
Breast
Breast
Breast
Breast
ATCC
ATCC
ATCC
ATCC
HTB-25
HTB-27
HTB-30
CRL-1500
Stromal cell line
CCD-1065SK skin (breast)
ATCC
CCD-1068Sk skin (breast)
CCD-1069Sk skin (breast)
ATCC
ATCC
HMF
Hs343.T
PC87322A1
PC87399A1
CRL-2077
CRL-2086
CRL-2089
Breast - Mammary fibroblasts
Breast - Mammry CAFs
Breast - Mammry CAFs
Breast - Mammry CAFs
Primary culture, CAFs
Primary culture, CAFs
Primary culture, CAFs
Sciencell
ATCC
Asterand
Asterand
7630
CRL-7245
PC87322A1-CAF
PC87399A1-CAF
Primary culture
CAFs
Primary culture, CAFs
Primary culture, CAFs
환자의 cancer sample 의 heterogeneity 유지는?
Breast cancer cell Marker
Scientists have identified a group of surface markers on cells linked to an aggressive type of breast cancer called estrogen receptor-negative canc
er. In this preliminary study, estrogen-negative breast cancer developed when three markers, CD44+, CD49fhi, and CD133hi were present sim
ultaneously on the surface of human cells taken from breast cancer patients and transplanted into a mouse; this is called a xenograft model. The s
cientists named these human cells with tumor-forming ability in mice, xenograft-initiating cells, or XIC. The research, conducted by scientists at the
National Cancer Institute (NCI), part of the National Institutes of Health, appeared online May 18, 2010, and in print June 1, 2010, in Cancer Rese
arch.
National cancer institute http://www.cancer.gov/newscenter/newsfromnci/2010/TripleMarkerENbreastcancer
Breast cancer stem cell markers – the rocky road to clinical applications http://breast-cancer-research.com/content/10/5/110
Molecular Profiling of Breast Cancer Cell Lines Defines Relevant Tumor Models and Provide
s a Resource for Cancer Gene Discovery
2009 Plus one http://www.plosone.org/article/info:doi/10.1371/journal.pone.0006146?imageURI=info:doi/10.1371/journal.pone.0006146.t001
Significance, detection and markers of disseminated breast cancer cells
Endocrine-Related Cancer (2006) 13 1033–1067 http://erc.endocrinology-journals.org/content/13/4/1033.full
Breast cancer는 2가지 type로 나눌 수 있다.
Luminal, basal type.
Luminal type : CD44-/CD24+
Basal type : CD44+/CD24stem/progenitor breast cancer cells can give rise to the heterogeneity of differentiated cells
proliferation이 강하게 나타난다.
The ability to form mammospheres is a characteristic of cancer stem cells. Tumors are broadly classified
histopathologically by expression of either luminal cytokeratins (CK18/19) or basal cytokeratin (CK5),
alpha smooth muscle actin (SMA) and vimentin
Homogeneous marker : CD29 (beta1-integrin), CD55 (DAF), CD59 (MIRL), CD81
(TAPA1), CD151 (PETA-3), CD166 (ALCAM). CD55 (DAF), and CD59 (MIRL)
No expression or low expression : CD66b, CD66c, CD165, and CD324
mesenchymal stem cells : CD44 (H-CAM), CD54 (ICAM), CD90 (Thy-1), CD105 (Endoglin), CD184
(CXCR4), various integrin molecules, such as CD49b (α 2integrin), CD49f (α 6integrin), CD29 (β1integrin),
CD61 (β3integrin), and other adhesion molecules including CD54 (ICAM) and CD166 (ALCAM)
myoepithelial/basal-like breast cancers (CK5, SMA, and vimentin)
luminal tumors (CK18 and CK19)
Breast cancer cell Marker
Breast cancer cell Marker
Significance, detection and markers of disseminated breast cancer cells
Endocrine-Related Cancer (2006) 13 1033–1067 http://erc.endocrinology-journals.org/content/13/4/1033.full
Breast cancer cell Marker
Breast cancer cell Marker
Breast cancer cell Marker
Breast cancer cell Marker
Breast cancer cell Marker
Primary Cell Culture into Cancer and Stromal Cell of Tissue
Protocol(수정 중)
1. 조직으로부터 4군데 이상 또는 절반 정도 sample을 채취한다.(sample 1g)
(sample 대조 : 환자명, 기록 번호)
2. Sample을 RPMI에 넣어 이동을 한다.
(RPMI, antibiotic
3. Use HBSS and Sample washing 3 time.
4. 멸균 된 칼로 조직을 분 가량 chopping 한다.
5. collagenase type 2를 시간 처리한다.(360U/ml)
6. c.f.g 를 RPM hr 로 돌려준다.
7. Use HBSS and Sample washing.
8. c.f.g 를 RPM hr 로 돌려준다.
9. RBC 제거를 위해 ficoll을 3:4 비율로 넣어준다.
10. c.f.g를 RPM hr 로 돌려준다.
11. Use HBSS and Sample washing.
12. DMEM을 넣어 37℃ 5% CO2 incubation 시킨다.
DMEM Gibco GIB-10569-010
collagenase from colstridium histolyticum type Ⅱ sigma C6885-1G
Ficoll-paque plus GE 10014768
Nature Volume: 487, Pages: 500–504 Date published:
(26 July 2012) DOI: doi:10.1038
nature11183 Received 07 November 2011 Accepted
04 May 2012 Published online 04 July 2012
Impact Factor :
36,2
NO . 0001
http://www.nature.com/nature/journal/v487/n7408/full/nature11183.html#/suppl
ementary-information
Tumour micro-environment elicits innate resistance to
RAF inhibitors through HGF secretion
Ravid Straussman,1 Teppei Morikawa,2 Kevin Shee,1 Michal Barzily-Rokni,1 Zhi Rong Qian,2 Jinyan Du,1 Ashli Davis,1 Margaret M. Mongare,1 Joshua G
ould,1 Dennie T. Frederick,3 Zachary A. Cooper,3 Paul B. Chapman,4 David B. Solit,4, 5 Antoni Ribas,6, 7 Roger S. Lo,7, 8 Keith T. Flaherty,3 Shuji Ogino,2,
9 Jennifer A. Wargo3 & Todd R. Golub1, 10, 11, 12
Fibroblast and cancer cell use to Co-culture, they are showed that
increase HGF from fibroblast.
Inhibition HGF signal (Raf, MET pathway), Melanoma and colorectal
cancer line reduce about proliferation.
Main actor : PLX4720(Raf inhibitor), PD184352(MEK inhibitor)
Many studies have focused on cell-autonomous mechan
isms of drug resistance.
Oncoprotein-targeted drugs hold enormous promise
for the future of cancer treatment.
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