Abstract
Cytological cancer screening that targets genetic or epigenetic abnormalities may be a viable alternative to morphological screening. Detecting cancer cells by specific genetic markers helps their easy detection in cytological samples. We recently established the telomerase-specific replication-selective adenovirus OBP-401, in which the human telomerase reverse transcriptase (hTERT) gene promoter has been inserted upstream of the E1 genes, and in which the green fluorescent protein (GFP) gene is driven by the CMV promoter. This virus selectively replicates only in telomerasepositive cells, expressing GFP, and therefore may be a tool for cancer screening. In the present study, we first confirmed that cytological samples can easily be infected with OBP-401, allowing visualization of GFP-positive cells under fluorescent microscopy 24 h after infection. After 32 cytological samples from patients with cervical, endometrial or ovarian cancers were infected with OBP-401, GFP signals were detected in 31 (96%) of the samples. However, some normal endometrial scrapings exhibited GFP-signals, possibly due to endometrial glandular cells with constitutive telomerase activity. The ability of OBP-401 to enrich cancer cells was then tested. Cytological samples containing cervical or endometrial cancer cells were infected with OBP-401, and GFP-positive cells were sorted by flow cytometry; DNA was extracted from the GFP-positive cells. Direct DNA sequencing or methylationspecific PCR identified cancer-derived mutations or hypermethylations of tumor suppressor genes more efficiently than analyses using crude cytological samples. Thus, OBP-401-based sorting of GFP-positive cells successfully enriched cancer cells, allowing efficient detection of genetic or epigenetic abnormalities in cytological samples.
| Original language | English |
|---|---|
| Pages (from-to) | 1549-1556 |
| Number of pages | 8 |
| Journal | International Journal of Oncology |
| Volume | 34 |
| Issue number | 6 |
| DOIs | |
| Publication status | Published - 2009 |
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Keywords
- Adenovirus
- Cytological screening
- Gynecologic cancers
- Molecular diagnosis
- Telomerase
ASJC Scopus subject areas
- Cancer Research
- Oncology
Cite this
Diagnostic potential and limitation of imaging cancer cells in cytological samples using telomerase-specific replicative adenovirus. / Maida, Yoshiko; Kyo, Satoru; Sakaguchi, Junko; Mizumoto, Yasunari; Hashimoto, Manabu; Mori, Noriko; Ikoma, Tomomi; Nakamura, Mitsuhiro; Takakura, Masahiro; Urata, Yasuo; Fujiwara, Toshiyoshi; Inoue, Masaki.
In: International Journal of Oncology, Vol. 34, No. 6, 2009, p. 1549-1556.Research output: Contribution to journal › Article
}
TY - JOUR
T1 - Diagnostic potential and limitation of imaging cancer cells in cytological samples using telomerase-specific replicative adenovirus
AU - Maida, Yoshiko
AU - Kyo, Satoru
AU - Sakaguchi, Junko
AU - Mizumoto, Yasunari
AU - Hashimoto, Manabu
AU - Mori, Noriko
AU - Ikoma, Tomomi
AU - Nakamura, Mitsuhiro
AU - Takakura, Masahiro
AU - Urata, Yasuo
AU - Fujiwara, Toshiyoshi
AU - Inoue, Masaki
PY - 2009
Y1 - 2009
N2 - Cytological cancer screening that targets genetic or epigenetic abnormalities may be a viable alternative to morphological screening. Detecting cancer cells by specific genetic markers helps their easy detection in cytological samples. We recently established the telomerase-specific replication-selective adenovirus OBP-401, in which the human telomerase reverse transcriptase (hTERT) gene promoter has been inserted upstream of the E1 genes, and in which the green fluorescent protein (GFP) gene is driven by the CMV promoter. This virus selectively replicates only in telomerasepositive cells, expressing GFP, and therefore may be a tool for cancer screening. In the present study, we first confirmed that cytological samples can easily be infected with OBP-401, allowing visualization of GFP-positive cells under fluorescent microscopy 24 h after infection. After 32 cytological samples from patients with cervical, endometrial or ovarian cancers were infected with OBP-401, GFP signals were detected in 31 (96%) of the samples. However, some normal endometrial scrapings exhibited GFP-signals, possibly due to endometrial glandular cells with constitutive telomerase activity. The ability of OBP-401 to enrich cancer cells was then tested. Cytological samples containing cervical or endometrial cancer cells were infected with OBP-401, and GFP-positive cells were sorted by flow cytometry; DNA was extracted from the GFP-positive cells. Direct DNA sequencing or methylationspecific PCR identified cancer-derived mutations or hypermethylations of tumor suppressor genes more efficiently than analyses using crude cytological samples. Thus, OBP-401-based sorting of GFP-positive cells successfully enriched cancer cells, allowing efficient detection of genetic or epigenetic abnormalities in cytological samples.
AB - Cytological cancer screening that targets genetic or epigenetic abnormalities may be a viable alternative to morphological screening. Detecting cancer cells by specific genetic markers helps their easy detection in cytological samples. We recently established the telomerase-specific replication-selective adenovirus OBP-401, in which the human telomerase reverse transcriptase (hTERT) gene promoter has been inserted upstream of the E1 genes, and in which the green fluorescent protein (GFP) gene is driven by the CMV promoter. This virus selectively replicates only in telomerasepositive cells, expressing GFP, and therefore may be a tool for cancer screening. In the present study, we first confirmed that cytological samples can easily be infected with OBP-401, allowing visualization of GFP-positive cells under fluorescent microscopy 24 h after infection. After 32 cytological samples from patients with cervical, endometrial or ovarian cancers were infected with OBP-401, GFP signals were detected in 31 (96%) of the samples. However, some normal endometrial scrapings exhibited GFP-signals, possibly due to endometrial glandular cells with constitutive telomerase activity. The ability of OBP-401 to enrich cancer cells was then tested. Cytological samples containing cervical or endometrial cancer cells were infected with OBP-401, and GFP-positive cells were sorted by flow cytometry; DNA was extracted from the GFP-positive cells. Direct DNA sequencing or methylationspecific PCR identified cancer-derived mutations or hypermethylations of tumor suppressor genes more efficiently than analyses using crude cytological samples. Thus, OBP-401-based sorting of GFP-positive cells successfully enriched cancer cells, allowing efficient detection of genetic or epigenetic abnormalities in cytological samples.
KW - Adenovirus
KW - Cytological screening
KW - Gynecologic cancers
KW - Molecular diagnosis
KW - Telomerase
UR - http://www.scopus.com/inward/record.url?scp=67649429234&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=67649429234&partnerID=8YFLogxK
U2 - 10.3892/ijo_00000284
DO - 10.3892/ijo_00000284
M3 - Article
C2 - 19424572
AN - SCOPUS:67649429234
VL - 34
SP - 1549
EP - 1556
JO - International Journal of Oncology
JF - International Journal of Oncology
SN - 1019-6439
IS - 6
ER -