Detection of circulating tumor cells in patients with gastrointestinal tract cancer using RT-PCR and its clinical implications

Noh, Yun Hee; Kim, Jung Ah; Lim, Grewo; Ro, Young Tae; Koo, Ja Hyun; Lee, Yong Sung; Han, Dong Soo; Park, Hwon Kyum; Ahn, Myung Ju

doi:10.1038/emm.2001.2

Cited by 18 publications

(9 citation statements)

References 24 publications

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“…In the literature, the presence of CTCs in the peripheral blood has been reported to be associated with malignant biological properties, more advanced disease and poor prognosis of GC patients. [12][13][14]30 Similarly, our results showed that the frequency of hTERT, CK-19, CEA and MUC1 mRNA overexpression were significantly higher in advanced GC patients (82.8-87.9%) than in patients with early stage GC (33.3-50%). Moreover, the detection rates for CTCs in GC patients by using single markers CK-19, CEA or MUC1 mRNA were prominently higher in GC patients with stage III and IV GC than in patients with stage I and II GC.…”

Section: Discussionsupporting

confidence: 53%

“…To date, several tumor-associated mRNA markers have been demonstrated to be absent in normal cells but overexpressed in GC cells, for example, human telomerase reverse transcriptase (hTERT), cytrokeratin-19 (CK-19), carcinoembryonic antigen (CEA) and mucin 1 (MUC1). [12][13][14][15][16] These mRNA markers have been extensively tested for their application to the detection of circulating tumor cells (CTCs) in GC patients.…”

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confidence: 99%

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Development of a high‐throughput membrane‐array method for molecular diagnosis of circulating tumor cells in patients with gastric cancers

Lin

et al. 2006

Intl Journal of Cancer

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Recently several noninvasive methods have been employed to detect circulating tumor cells (CTCs) in cancer patients. In this study, we have developed a highly sensitive, high-throughput colorimetric membrane-array method that was designed to detect a panel of mRNA markers including human telomerase reverse transcriptase (hTERT), , carcinoembryonic antigen (CEA) and mucin 1 (MUC1) mRNA for the presence of CTCs in the peripheral blood of patients with gastric cancer (GC). Digoxigenin-labeled cDNA targets synthesized following total RNA isolation from peripheral blood samples of 64 GC patients and 80 healthy individuals were subjected to membrane-array hybridization. The results showed that membrane array could positively detect 5 cancer cells/ml of peripheral blood in GC cell-dilution experiments. The sensitivity, specificity and diagnostic accuracy for hTERT, CK-19, CEA and MUC1 mRNA ranged from 78.1% to 82.8%, 76.3% to 85% and 81.3% to 83.3%, respectively. Both CEA and MUC1 mRNA expression was correlated significantly with all malignant biological properties of GC, such as macroscopic type, depth of tumor invasion, lymph-node metastasis, TNM stage and coexisting distant metastasis (all p < 0.05). Using these 4 markers in combination, the sensitivity, specificity and diagnostic accuracy of membrane array were raised to 89.1%, 91.3% and 90.3%, respectively. The expression of all 4 mRNA markers was an independent predictor for postoperative recurrence/metastasis. GC patients with the expression of all the 4 mRNA markers showed a poorer survival rate than those without the expression of any 1 mRNA marker (p 5 0.0223). These findings demonstrated that our membrane-array method could detect CTCs in the circulation of GC patients with considerably high sensitivity and specificity. The identification of CTCs in the peripheral blood may be useful in the auxiliary cancer diagnostics or postoperative surveillance of GC patients for recurrence/metastasis. ' 2006 Wiley-Liss, Inc.Key words: circulating tumor cell; membrane array; gastric cancer; molecular diagnosisThe degrees of tumor penetration in the stomach wall and lymph node metastasis have been used for many decades as the 2 major prognostic determinants for gastric cancer (GC) patients. Unfortunately, despite informative staging of patients with GC, some patients with apparently localized disease at diagnosis will subsequently develop recurrent or metastatic diseases. 1-4 One of the major causes is attributed to the presence of disseminated tumor cells shed from the primary carcinoma into circulation, prior to or during surgery. Even in patients with early GC, blood-born metastasis occurs. 2,5,6 Recent attempts to improve staging include sensitive detection of disseminated tumor cells in the blood or lymph nodes by molecular approaches. Evidence increasingly clarifies that primary cancers begin shedding neoplastic cells into the circulation at an early stage. [7][8][9] It is the dissemination of cancer cells into circulation that is widely accepted as the main cause l...

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Section: Discussionsupporting

confidence: 53%

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confidence: 99%

Development of a high‐throughput membrane‐array method for molecular diagnosis of circulating tumor cells in patients with gastric cancers

Lin

et al. 2006

Intl Journal of Cancer

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“…To confirm the differential expressions of genes screen-ed by DNA chip analysis, RT-PCR was performed (Noh et al, 2001). To synthesize first strand cDNA, one microgram of RNA and 1 µl of 10 µM oligodT (T25NN) were mixed to a final reaction volume of 5 µl and heated for 2 min at 72 o C. After cooling on ice for 2 min, RT was performed for 2 h at 42 o C in a 10 µl reaction mixture containing 50 mM Tris-HCl, pH 8.3, 75 mM KCl, 6 mM MgCl2, 2 mM DTT, 1 mM dNTP mix and 200 units MMLV reverse transcriptase.…”

Section: Reverse Transcription-polym Erase Chain Reaction (Rt-pcr)mentioning

confidence: 99%

Identification of amyloid β-peptide responsive genes by cDNA microarray technology: Involvement of RTP801 in amyloid β-peptide toxicity

Kim

Lee²,

Chung³

et al. 2003

Exp Mol Med

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Abbreviations: Aβ, Amyloid β-peptide; AD, Alzheimer's disease; CDK, cyclin-dependent kinase; DMEM, Dulbecco's modified Eagle's medium; DSCR1, Down syndrome candidate region 1; MTT, 3-(4, 5-dimethylthiazol-2-yl)-2, 5-diphenyltetrazolium bromide; RT-PCR, reverse transcription polymerase chain reaction; SDS, sodium dodecylsulfate; SEST2, Hi95/sestrin 2; STC, stanniocalcin; ZFP36L2, zinc finger protein 36. A bstract

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“…Total RNA was isolated from the human spermatozoa using the guanidinium thiocyanate-phenol extraction (Chomczynski and Sacchi, 1987;Noh et al, 2001), which was repeated at least 3 times to improve the quality of sperm RNA.…”

Section: Aterials and M Ethodsmentioning

confidence: 99%

Molecular identification of Ca(2+)channels in human sperm

Park

Ahn²,

Gu³

et al. 2003

Exp Mol Med

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A bstractThe acrosom e reaction is a Ca 2+ -dependent exocytotic process that is a prerequisite step for fertilization. External calcium entry through voltage-activated Ca 2+ channels is know n to be essential in inducing the acrosome reaction of mammalian sperm atozoa. Due to their com plex geom etry, however, electrophysiological identification of sperm Ca 2+ channels has been lim ited. Here we identified Ca 2+ channel m RNAs expressed in m otile hum an sperm using RT-PCR and their levels w ere compared using RNase protection assays. L-type, non-L-type, and T-type C a 2+ channel m RNAs w ere detected by RT-PCR using degenerate prim ers. Cloning and sequencing of the PCR products revealed α1B, α1C, α1E, α1G , and α1H sequences. RT-PCR using specific prim ers repeatedly detected α1B, α1C, α1E, α1G , and α1H m RNAs, and additionally α1I m RNA. But α1A and α1D m essages w ere not detected. Relative expression levels of the detected Ca 2+ channel subtypes were compared by RNase protection assays. The abundance of detected m RNA m essages w as in the follow ing order: α1H α1G α1E α1B α1C α1I. These findings indicated that hum an m otile sperm express m ultiple voltage-activated Ca 2+ channel RNAs am ong which T-type and non-L-type channel m essages are likely to be predom inantly expressed. Based on their relative expression levels, w e propose that not only T-type but also non-L-type calcium channels m ay be m ajor gates for the external calcium influx, required for the acrosom e reaction.

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Detection of circulating tumor cells in patients with gastrointestinal tract cancer using RT-PCR and its clinical implications

Cited by 18 publications

References 24 publications

Development of a high‐throughput membrane‐array method for molecular diagnosis of circulating tumor cells in patients with gastric cancers

Development of a high‐throughput membrane‐array method for molecular diagnosis of circulating tumor cells in patients with gastric cancers

Identification of amyloid β-peptide responsive genes by cDNA microarray technology: Involvement of RTP801 in amyloid β-peptide toxicity

Molecular identification of Ca(2+)channels in human sperm

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