scholarly journals Comparison of Gene Expression in Breast Cancer Using Laser Capture Microdissection and Quantitative RT-PCR

2003 ◽  
Vol 36 (5) ◽  
pp. 421-425 ◽  
Author(s):  
Xiaojuan Wang ◽  
Misa Nakamura ◽  
Ichiro Mori ◽  
Koichi Takeda ◽  
Takaomi Suzuma ◽  
...  
Keyword(s):  
Breast Cancer ◽  
Gene Expression ◽  
Laser Capture Microdissection ◽  
Rt Pcr ◽  
Laser Capture

410. Gene expression analysis in endometriotic lesions using laser capture microdissection

2008 ◽  
Vol 20 (9) ◽  
pp. 90
Author(s):  
L. Fu ◽  
J. E. Girling ◽  
P. A. W. Rogers
Keyword(s):  
Gene Expression ◽  
Real Time ◽  
Laser Capture Microdissection ◽  
Stromal Cells ◽  
Expression Profiles ◽  
Specific Gene ◽  
Rt Pcr ◽  
Normal Endometrium ◽  
Rna Quality ◽  
Laser Capture

Previous studies examining gene expression profiles in normal endometrium and endometriotic lesions have used RNA extracted from whole tissue samples. Results from these studies can be difficult to interpret as they reflect expression averaged across several different cell types that may be functionally quite different. The aim of this study was to establish laser capture microdissection (LCM) as a technique to examine gene expression in stromal and epithelial cells from normal and ectopic endometrium. We hypothesised that genes associated with inflammation would be elevated in cells from endometriotic lesions. Full thickness uterine samples were collected during abdominal hysterectomy from normal cycling premenopausal women. Endometriotic lesions were collected during abdominal laparoscopy. Samples were either frozen in OCT or stored in RNAlater for 12 h before freezing. Tissues were immunostained with an antibody against CD10 to identify ectopic endometrial stromal cells before LCM. Endometrial epithelial and stromal cells were collected using the PALM MicroLaser System. RNA quality was accessed using Experion. TGFβ1, MMP1, αSMA, SMAD2 and NFκB mRNA was analysed using real-time RT–PCR. Of the endometriotic samples stored in OCT (n = 58), only 14% (n = 8) had visible endometrial glands. Of these, only 37% (n = 3) had RNA of an acceptable quality for further analysis. However, RNA quality and quantity were dramatically improved in 3 of 5 samples collected in RNAlater. In preliminary studies, expression of TGFβ1 and αSMA mRNA was elevated in endometriotic lesions in comparison to the normal endometrium, whereas NFκB expression did not change. We have shown that RNAlater solution is useful to preserve RNA quality for small clinical endometriotic samples and that immuno-guided LCM-generated homogenous cell populations coupled with real-time RT–PCR can provide valuable insights into cell and disease-specific gene expression in endometriotic lesions.

scholarly journals Analysis of gene expression in mineralized skeletal tissues by laser capture microdissection and RT-PCR

2006 ◽  
Vol 86 (10) ◽  
pp. 1089-1095 ◽  
Author(s):  
Yvonne Y Shao ◽  
Lai Wang ◽  
David G Hicks ◽  
R Tracy Ballock
Keyword(s):  
Gene Expression ◽  
Laser Capture Microdissection ◽  
Rt Pcr ◽  
Laser Capture

scholarly journals TRIGONELLA FOENUM-GRAECUM L. EXHIBITS ESTROGENIC EFFECT THROUGH PRESENILIN 2 GENE EXPRESSION IN THE BREAST CANCER CELL LINE MCF-7

2019 ◽  
Vol 12 (1) ◽  
pp. 438
Author(s):  
Kurnia Agustini ◽  
Michael Wink ◽  
Wahono Sumaryono ◽  
Frans Suyatna ◽  
Nurjati Chairani Siregar
Keyword(s):  
Breast Cancer ◽  
Gene Expression ◽  
Cancer Cell Line ◽  
Active Fraction ◽  
Rt Pcr ◽  
Fenugreek Seeds ◽  
Trigonella Foenum Graecum ◽  
Presenilin 2 ◽  
Trigonella Foenum Graecum L ◽  
Mcf 7

Objective: The objective of this study is to investigate the estrogenic and antiestrogenic activity of Fenugreek seeds, Trigonella foenum-graecum L. in the estrogen-dependent breast cancer cell line, MCF-7, including its effect on the expression of estrogen-dependent presenilin 2 (pS2) gene.Methods: An activity guided fractionation was carried out with extracts from fenugreek seeds in MCF-7 cells. Cytotoxic activity assays were conducted with 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium (MTT) assay. Most fractions were also tested also tested in media with estradiol 10 nM We also analysed the expression of pS2 gene. For the analysis of pS2 gene expression we employed PCR primers for pS2 and for β-actin as a housekeeping gene using real-time polymerase chain reaction (RT-PCR).Results: Based on cytotoxic activity assay in MCF-7, the active fractions are ethyl acetic fraction and its phases ethyl acetic (EA) 2 and EA 2.2. The most active fraction was EA 2.2 (IC50=27.129 ppm), which exhibited a biphasic effect; at low concentrations, it stimulated the growth, and at high concentrations it showed strong cytotoxic effects. EA2.2 fraction in concentration 20 ppm, also could induce pS2 gene expression in media with and without estrogen.Conclusion: The most active fraction was the ethyl acetate phase and further subfractions. The most active fraction also induced the expression of pS2 gene which was studied by RT-PCR.

Sign in / Sign up

Export Citation Format

Share Document