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Identification of molecular markers for the early detection of human squamous cell carcinoma of the uterine cervix.

Cheng Q, Lau WM, Chew SH, Ho TH, Tay SK, Hui KM - Br. J. Cancer (2002)

Bottom Line: Of particular interest is clone G30CC that has been identified to be the gene that encodes S12 ribosomal protein.When employed for RNA--RNA in situ hybridization experiments, expression of G30CC could be detected in the immature basal epithelial cells of histo-pathological normal tissues collected from cervical cancer patients of early FIGO stages.In comparison, the expression of G30CC was not detected in cervical tissues collected from patients admitted for surgery of non-malignant conditions.

View Article: PubMed Central - PubMed

Affiliation: Laboratory of Gene Structure & Expression, Division of Cellular and Molecular Research, National Cancer Centre, 11 Hospital Drive, 169610 Singapore.

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Related in: MedlinePlus

Differential display analysis. Purified total RNA from cervical cancer biopsies was reversed transcribed with the primer P21 (see Materials and Methods). The reverse transcribed cDNA were then amplified by PCR in the presence of 1 μCi [α-33P]dATP and either primer P30, P31, or P32. The PCR products were then separated on polyacrylamide gel. (A) RT–PCR differential display with the primer pair P21/P30. (B) RT–PCR differential display with the primer pair P21/P31. (C) RT–PCR differential display with the primer pair P21/P32. Seventeen bands selected following differential display analyses with the corresponding primer pairs, as indicated in the figure, were employed for subsequent cloning.
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fig1: Differential display analysis. Purified total RNA from cervical cancer biopsies was reversed transcribed with the primer P21 (see Materials and Methods). The reverse transcribed cDNA were then amplified by PCR in the presence of 1 μCi [α-33P]dATP and either primer P30, P31, or P32. The PCR products were then separated on polyacrylamide gel. (A) RT–PCR differential display with the primer pair P21/P30. (B) RT–PCR differential display with the primer pair P21/P31. (C) RT–PCR differential display with the primer pair P21/P32. Seventeen bands selected following differential display analyses with the corresponding primer pairs, as indicated in the figure, were employed for subsequent cloning.

Mentions: One microgram of total RNA extracted from each cervical biopsy was reverse transcribed using the P21 primer. This was followed by a subsequent PCR amplification step employing the P21/P30, P21/P31, or P21/P32 PCR primer pairs. All PCR reactions, including the RT step, were performed in triplicates to ensure reproducibility. In addition, each of the PCR products was analyzed in different electrophoretic separations employing denaturing polyacrylamide gels with various running times. The gel patterns of the amplified PCR products of six independent mRNAs purified from different cancer biopsies and matched normal tissues gave virtually identical gel patterns for each of the PCR primer set employed (Figure 1Figure 1


Identification of molecular markers for the early detection of human squamous cell carcinoma of the uterine cervix.

Cheng Q, Lau WM, Chew SH, Ho TH, Tay SK, Hui KM - Br. J. Cancer (2002)

Differential display analysis. Purified total RNA from cervical cancer biopsies was reversed transcribed with the primer P21 (see Materials and Methods). The reverse transcribed cDNA were then amplified by PCR in the presence of 1 μCi [α-33P]dATP and either primer P30, P31, or P32. The PCR products were then separated on polyacrylamide gel. (A) RT–PCR differential display with the primer pair P21/P30. (B) RT–PCR differential display with the primer pair P21/P31. (C) RT–PCR differential display with the primer pair P21/P32. Seventeen bands selected following differential display analyses with the corresponding primer pairs, as indicated in the figure, were employed for subsequent cloning.
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Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC2375172&req=5

fig1: Differential display analysis. Purified total RNA from cervical cancer biopsies was reversed transcribed with the primer P21 (see Materials and Methods). The reverse transcribed cDNA were then amplified by PCR in the presence of 1 μCi [α-33P]dATP and either primer P30, P31, or P32. The PCR products were then separated on polyacrylamide gel. (A) RT–PCR differential display with the primer pair P21/P30. (B) RT–PCR differential display with the primer pair P21/P31. (C) RT–PCR differential display with the primer pair P21/P32. Seventeen bands selected following differential display analyses with the corresponding primer pairs, as indicated in the figure, were employed for subsequent cloning.
Mentions: One microgram of total RNA extracted from each cervical biopsy was reverse transcribed using the P21 primer. This was followed by a subsequent PCR amplification step employing the P21/P30, P21/P31, or P21/P32 PCR primer pairs. All PCR reactions, including the RT step, were performed in triplicates to ensure reproducibility. In addition, each of the PCR products was analyzed in different electrophoretic separations employing denaturing polyacrylamide gels with various running times. The gel patterns of the amplified PCR products of six independent mRNAs purified from different cancer biopsies and matched normal tissues gave virtually identical gel patterns for each of the PCR primer set employed (Figure 1Figure 1

Bottom Line: Of particular interest is clone G30CC that has been identified to be the gene that encodes S12 ribosomal protein.When employed for RNA--RNA in situ hybridization experiments, expression of G30CC could be detected in the immature basal epithelial cells of histo-pathological normal tissues collected from cervical cancer patients of early FIGO stages.In comparison, the expression of G30CC was not detected in cervical tissues collected from patients admitted for surgery of non-malignant conditions.

View Article: PubMed Central - PubMed

Affiliation: Laboratory of Gene Structure & Expression, Division of Cellular and Molecular Research, National Cancer Centre, 11 Hospital Drive, 169610 Singapore.

Show MeSH
Related in: MedlinePlus