Skip to main content
Key Specialties
Cardiology Diabetology Endocrinology Gastroenterology Geriatrics and Gerontology Gynecology and Obstetrics Hematology Infectious Disease Internal Medicine Nephrology Neurology Oncology Pediatrics Respiratory Medicine Rheumatology Surgery
Congress Coverage
2024 ACC Congress 2023 ESMO Congress 2023 EASD Congress 2023 ERS Congress 2023 ESC Congress
Clinical Collections
Advances in Alzheimer’s

Keynote webinar | Spotlight on medication adherence

LIVE: Thursday 27th June 2024, 18:00-19:30 (CEST)
Join our expert panel to discover why you need to understand the drivers of non-adherence in your patients, and how you can optimize medication adherence in your clinics to drastically improve patient outcomes.
ADVANCED SEARCH
Log in
Top
BMC Cancer
Published in: BMC Cancer 1/2008

Open Access 01-12-2008 | Research article

Efficacy of laser capture microdissection plus RT-PCR technique in analyzing gene expression levels in human gastric cancer and colon cancer

Authors: Hiroshi Makino, Hiroyuki Uetake, Kathleen Danenberg, Peter V Danenberg, Kenichi Sugihara

Published in: BMC Cancer | Issue 1/2008

Login to get access

Abstract

Background

Thymidylate synthase, dihydropyrimidine dehydrogenase, thymidine phosphorylase, and orotate phosphoribosyltransferase gene expressions are reported to be valid predictive markers for 5-fluorouracil sensitivity to gastrointestinal cancer. For more reliable predictability, their expressions in cancer cells and stromal cells in the cancerous tissue (cancerous stroma) have been separately investigated using laser capture microdissection.

Methods

Thymidylate synthase, dihydropyrimidine dehydrogenase, thymidine phosphorylase, and orotate phosphoribosyltransferase mRNA in cancer cells and cancerous stroma from samples of 47 gastric and 43 colon cancers were separately quantified by reverse transcription polymerase chain reaction after laser capture microdissection.

Results

In both gastric and colon cancers, thymidylate synthase and orotate phosphoribosyltransferase mRNA expressions were higher (p < 0.0001, p <0.0001 respectively in gastric cancer and P = 0.0002, p < 0.0001 respectively in colon cancer) and dihydropyrimidine dehydrogenase mRNA expressions were lower in cancer cells than in cancerous stroma (P = 0.0136 in gastric cancer and p < 0.0001 in colon cancer). In contrast, thymidine phosphorylase mRNA was higher in cancer cells than in cancerous stroma in gastric cancer (p < 0.0001) and lower in cancer cells than in cancerous stroma in colon cancer (P = 0.0055).

Conclusion

By using this method, we could estimate gene expressions separately in cancer cells and stromal cells from colon and gastric cancers, in spite of the amount of stromal tissue. Our method is thought to be useful for accurately evaluating intratumoral gene expressions.
Appendix
Available only for authorised users
Literature
1.
Jemal A, Murray T, Ward E, Samuels A, Tiwari RC, Ghafoor A, Feuer EJ, Thun MJ: Cancer Statistics, 2005. CA Cancer J Clin. 2005, 55: 10-30.CrossRefPubMed
2.
Ajamni JA: Evolving chemotherapy for advanced gastric cancer. Oncologist. 2005, 10 Suppl 3: 49-58.CrossRef
3.
Saltz LB, Cox JV, Blanke C: Irinotecanplus fluorouracil and leucovorin for metastatic colorectal cancer. Irinotecan Study Group. N Engl J Med. 2000, 343: 905-914.CrossRefPubMed
4.
Douillard JY, Cunningham D, Roth AD: Iriniotecan combined with fluorouracil compared with fluorouracil alone as first-line treatment for metastatic colorectal cancer: A multicentre randomized trial. Lancet. 2000, 355: 1041-1047.CrossRefPubMed
5.
Goldberg RM, Sargent DJ, Morton RF: A randomized controlled trial of fluorouracil plus leucovorin, irinotecan, and oxaliplatin combinations in patients with previously untreated metastatic colorectal cancer. J Clin Oncol. 2004, 22: 23-30.CrossRefPubMed
6.
de Gramont A, Figer A, Seymour M: Leucovorin and fluorouracil with or without oxaliplatin as first-line treatment in advanced colorectal cancer. J Clin Oncol. 2000, 18: 2938-2947.PubMed
7.
Grothey A, Deschler B, Kroening H: Phase III study of bolus 5-fluorouracil (5-FU)/folinic acid (FA) (Mayo) vs weekly high-dose 24 h 5-FU infusion/FA + oxaliplatin in advanced colorectalcancer. Proc Am Soc Clin Oncol. 2002, 21: 129a-(abstr 512)
8.
Ohtsu A: Current status and future prospects of chemotherapy for metastatis gastric cancer:a review. Gastric Cancer. 2005, 8: 95-102.CrossRefPubMed
9.
Danenberg PV: Thymidylate synthase – a target enzyme in cancer chemotherapy. Biochem Biophys Acta. 1977, 473: 73-92.PubMed
10.
Peters GJ, Wilt van der CL, van Triest B, Codacci-Pisanelli G, Johnston PG, van Groeningen CJ, Pinedo HM: Thymidylate syntase and drug resistance. Eur J Cancer. 1995, 31A: 1299-1305.CrossRefPubMed
11.
Salonga D, Danenberg KD, Johnson M, Metzger R, Groshen S, Tsao-Wei DD, Lenz HJ, Leichman L, Diasio RB, Danenberg PV: Colorectal tumors responding to 5-fluorouracil have low gene expression levels of dihydropyrimidine dehydrogenase, thymidylate syntase, and thymidine phosphorylase. Clin Cancer Res. 2000, 6: 1322-1327.PubMed
12.
Ichikawa W, Uetake H, Shirota Y, Yamada H, Nishi N, Nihei Z, Sugihara K, Hirayama R: Combination of dihydropyrimidine dehydrogenase and thymidylate syntase gene expressions in primary tumors as predictive parameters for the efficacy of fluoropyrimidine-based chemotherapy for metastatic colorectal cancer. Clin Cancer Res. 2003, 9: 786-789.PubMed
13.
Peters GJ, Laurensse E, Leyva A, Lankelma J, Pinedo HM: Sensitivity of human, murine, and rat cells to 5-fluorouraci and 5'-deoxy-5-fluorouracil in relation to drug-metabolizing enzymes. Caner Res. 1986, 46: 20-28.
14.
Koizumi W, Saigenji K, Nakamaru N, Okayasu I, Kurihara M: Prediction of response to 5'-deoxy-5-fluorouridine (5'-DFUR) in patients with inoperable advanced gastric cancer by immunostaining of thymidine phosphorylase/platelet-derived endotherial cell growth factor. Oncology. 1999, 56: 215-222.CrossRefPubMed
15.
Sawada N, Ishikawa T, Fukase Y, Nishida M, Yoshikubo T, Ishitsuka H: Induction of thymidine phosphorylase activity and enhancement of capecitabine efficacy by taxil/taxotere in human cancer xenografts. Clin Cancer Res. 1998, 4: 1013-1019.PubMed
16.
Peters GJ, Braakhuis BJ, Bruijin EA, Laurensse EJ, Walsum M, Pinedo HM: Enhanced therapeutic efficacy of 5'-deoxy – 5-fluorouridine in 5-fluorouracil-ressistant head and neck tumours in relation to 5-fluorouracil metabolizing enzymes. Br J Cancer. 1989, 59: 327-334.CrossRefPubMedPubMedCentral
17.
Ma T, Zhu ZC, Ji YB, Zhang Y, Yu YY, Liu BY, Yin HR, Lin YZ: Correlation of thymidylate synthase, thymidine phosphorylase and dihydropyrimidine dehydrogenase with sensitivity of gastrointestinal cancer cells to 5-fluorouracil and 5-fluoro-2'-deoxyuridine. World J Gastroenterol. 2004, 10: 172-176.PubMedPubMedCentral
18.
Toriumi F, Kubota T, Saikawa Y, Yoshida M, Otani Y, Watanabe M, Kumai K, Kitajima M: Thymidylate synthase(TS)genotype and TS/dihydropyrimidine dehydrogenase mRNA levels as am indicator in determining chemosensitivity to 5-fluorouracil in advanced gastric carcinoma. Anticancer Res. 2004, 24: 2455-2463.PubMed
19.
Ishikawa Y, Kubota T, Otani Y, Watanabe M, Teramoto T, Kumai K, Takechi T, Okabe H, Fukushima M, Kitajima M: Dihydropyrimidine dehydrogenase and messenger RNA levels in gastric cancer possible predictor for sensetivity to 5-fluorouracil. Jpn J Cancer Res. 2000, 91: 105-112.CrossRefPubMed
20.
Ichikawa W, Uetake H, Shirota Y, Yamada H, Takahashi T, Nihei Z, Sugihara K, Sasaki Y, Hirayama R: Both gene expression for orotate phosphoribosyl transferase and its ratio to dihydropyrimidine dehydrogenase influence outcome following fluoropyrimidine-based chemotherapy for metastatic colorectal cancer. Br J Cancer. 2003, 89: 1486-1492.CrossRefPubMedPubMedCentral
21.
Ochiai T, Nishimura K, Noguchi H, Kitajima M, Tsukada A, Watanabe E, Nagaoka I, Futagawa S: Prognostic impact of orotate phosphoribosyl transferase among 5-fluorouracil metabolic enzymes ie respectable colorectal cancers treated by oral 5-fluorouracil based adjuvant chemotherapy. Int J Cancer. 2006, 118: 3084-3088.CrossRefPubMed
22.
Adlard JW, Richman SD, Seymour MT, Quirke P: Prediction of the response of colorectal cancer to systemic therapy. Lancet Oncol. 2002, 3: 75-82.CrossRefPubMed
23.
Isshi K, Sakuyama T, Gen T, Nakamura Y, Kuroda T, Katsuyama T, Maekawa Y: Predicting 5-FU sensitivity using human colorectal cancer specimens: comparison of tumor dihydropyrimidine dehydrogenase and orotate phosphoribosyl transferase activities with in vitro chemosensitivity to 5-FU. Int J Clin Oncol. 2002, 7: 335-342.CrossRefPubMed
24.
Kakimoto M, Uetake H, Osanai T, Shirota Y, Sugihara K: Thymidine synthasae and dihydropyrimidine dehydrogenase gene expression in breast cancer predicts 5-FU sensitivity by a histocultural drug sensitivity test. Cancer Letters. 2005, 223: 103-111.CrossRefPubMed
25.
Lord RV, Salonga D, Danenberg KD, Peters JH, DeMeester TR, Park JM, Johansson J, Skinner KA, Chandrasoma P, DeMeester SR, Bremner CG, Tsai PI, Danenberg PV: Teromerase reverse transcriptase, expression in increased early in the Barrett's metaplasia, dysplasia, carcinoma sequence. J Gastrointest Surg. 2000, 4: 135-142.CrossRefPubMed
26.
Heid CA, Stevens J, Livak KJ, Williams PM: Real time quantitative RT-PCR. Gemome Res. 1996, 10: 986-994.
27.
Gibson UE, Heid CA, Williams PM: A novel method for real time quantitative RT-PCR. Gemome Res. 1996, 10: 995-1011.
28.
Sakamoto K, Sugimoto Y, Miyadera K, Oka T, Fukushima M: Preparation of anti-orotate phosphoribosyltransferase antibody and its application to immunohistochemical detection in human tumor cells. Int J Mol Med. 2005, 16: 245-249.PubMed
29.
Oguri T, Achiwa H, Bessho Y, Muramatsu H, Maeda H, Niimi T, Sato S, Ueda R: The role of thymidylate synthase and dihydropyrimidine dehydrogenase in resistance to 5-fluorouracil in human lung cancer cells. Lung Cancer. 2005, 49: 345-351.CrossRefPubMed
30.
Kamoshida S, Shiogama K, Shimomura R, Inada K, Sakurai Y, Ochiai M, Matuoka H, Maeda K, Tsutsumi Y: Immunohistochemical demonstration of fluoropyrimidine-metabolizing enzymes in various types of cancer. Oncol Rep. 2005, 14: 1223-1230.PubMed
31.
Peters G, Groeningen C, Laurensse E, Pinedo HM: A Comparison of 5-fluorouracil metabolism in human colorectal cancer and colon mucosa. Cancer. 1991, 68: 1903-1909.CrossRefPubMed
32.
Maehara Y, Moriguchi S, Emi Y, Watanabe A, Kohnoe S, Tsujitani S, Sugimachi K: Comparison of pyrimidine nucleotide synthetic enzymes involved in 5-fluoroyracil metabolism between human adnocarcinomas and squamous cell carcinomas. Cancer. 1990, 66: 156-161.CrossRefPubMed
33.
Ichikawa W, Takahashi T, Suto K, Nihei Z, Shirota Y, Shimizu M, Sasaki Y, Hirayama R: Thymidylate synthase and dihydropyrimidine dehydrogenase gene expression in relation to differentiation of gastric cancer. Int J Cancer. 2004, 112: 967-973.CrossRefPubMed
34.
Lassmann S, Hennig M, Rosenberg R, Nährig J, Schreglmann J, Krause F, Poignee-Heger M, Nekarda H, Höfler H, Werner M: Thymidine phosphorylase, dihydropyrimidine dehydrogenase and thymidylate synthase mRNA expression in primary colorectal tumors-correlation to tumor histopathology and clinical follow up. Int J Colorectal Dis. 2006, 21: 238-247.CrossRefPubMed
35.
Johnston S, Ridge S, Cassidy J, McLeod H: Reguration of dihydropyrimidine dehydrogenase in colorectal cancer. Clin Cancer Res. 1999, 5: 2566-2570.PubMed
36.
Takebayashi Y, Yamada K, Miyadera K, Sumizawa T, Furukawa T, Kinoshita F, Aoki D, Okumura H, Yamada Y, Akiyama S, Aikou T: The activity and expression of thymidine phosphorylase in human solid tumours. Eur J Cancer. 1996, 32A: 1227-1232.CrossRefPubMed
37.
Maeda K, Kang SM, Ogawa M, Onoda N, Sawada T, Nakata B, Kato Y, Chung YS, Sowa M: Combined analysis of vascular endothelial growth factor and platelet-derived endothelial cell growth factor expression in gastric carcinoma. Int J Cancer. 1997, 74: 545-550.CrossRefPubMed
38.
Toi M, Ueno T, Matsumoto H, Saji H, Funata N, Koike M, Tominaga T: Significance of thymidine phosphorylase as a marker of protumor monocytes in breast cancer. Clin Cancer Res. 1999, 5: 1131-1137.PubMed
39.
Takebayashi Y, Akiyama S, Akiba S, Yamada K, Miyadera K, Sumizawa T, Yamada Y, Murata F, Aikou T: Clinicopathologic and prognostic significance of an angiogenic factor, thymidine phosphorylase, in human colorectal carcinoma. J Natl Cancer Inst. 1996, 88: 1110-1117.CrossRefPubMed
40.
Amaya H, Tanigawa N, Lu C, Matsumura M, Shimomatsuya T, Horiuchi T, Muraoka R: Association of vascular endothelial growth factor expression with tumor angiogenesis, survival and thymidine phosphorylase/platelet-derived endothelial cell growth factor expression in human colorectal cancer. Cancer Lett. 1997, 119: 227-235.CrossRefPubMed
41.
Haba A, Monden T, Sekimoto M, Ikeda K, Izawa H, Kanou T, Amano M, Kanayama H, Monden M: PyNPase expression in human colon cancer. Cancer Lett. 1998, 122: 85-92.CrossRefPubMed
42.
Matsumura M, Chiba Y, Lu C, Amaya H, Shimomatsuya T, Horiuchi T, Muraoka R, Tanigawa N: Platelet-derived endothelial cell growth factor/thymidine phosphorylase expression correlated with tumor angiogenesis and macrophage infiltration in colorectal cancer. Cancer Lett. 1998, 128: 55-63.CrossRefPubMed
43.
Emmert-Buck MR, Bonner RF, Smith PD, Chuaqui RF, Zhuang Z, Godstein SR, Weiss RA, Liotta LA: Laser capture microdissection. Science. 1996, 274: 998-1001.CrossRefPubMed
44.
Farrugia DC, Ford HE, Cunningham D, Danenberg KD, Danenberg PV, Brabender J, McVicar AD, Aherne GW, Hardcastle A, McCarthy K, Jackman AL: Thymidylate synthase expression in advanced colo-rectal cancer predicts for response to raltitrexed. Clin Cancer Res. 2003, 9: 792-801.PubMed
Metadata
Title
Efficacy of laser capture microdissection plus RT-PCR technique in analyzing gene expression levels in human gastric cancer and colon cancer
Authors
Hiroshi Makino
Hiroyuki Uetake
Kathleen Danenberg
Peter V Danenberg
Kenichi Sugihara
Publication date
01-12-2008
Publisher
BioMed Central
Published in
BMC Cancer / Issue 1/2008
Electronic ISSN: 1471-2407
DOI
https://doi.org/10.1186/1471-2407-8-210

Other articles of this Issue 1/2008

BMC Cancer 1/2008 Go to the issue

Research article

Tumor necrosis is associated with increased alphavbeta3integrin expression and poor prognosis in nodular cutaneous melanomas

Research article

Enhancer of the rudimentary gene homologue (ERH) expression pattern in sporadic human breast cancer and normal breast tissue

Research article

Expression of HER-2 in MCF-7 breast cancer cells modulates anti-apoptotic proteins Survivin and Bcl-2 via the extracellular signal-related kinase (ERK) and phosphoinositide-3 kinase (PI3K) signalling pathways

Research article

Cervix carcinoma is associated with an up-regulation and nuclear localization of the dual-specificity protein phosphatase VHR

Research article

Retinoic acid reduces human neuroblastoma cell migration and invasiveness: effects on DCX, LIS1, neurofilaments-68 and vimentin expression

Research article

Characterization of three new serous epithelial ovarian cancer cell lines
Webinar | 19-02-2024 | 17:30 (CET)

Keynote webinar | Spotlight on antibody–drug conjugates in cancer

Watch now

Antibody–drug conjugates (ADCs) are novel agents that have shown promise across multiple tumor types. Explore the current landscape of ADCs in breast and lung cancer with our experts, and gain insights into the mechanism of action, key clinical trials data, existing challenges, and future directions.

Dr. Véronique Diéras
Prof. Fabrice Barlesi
Developed by: Springer Medicine
Image Credits