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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

Reverse Northern blot. DNA (2 μg) of the amplified gene fragment of each of the purified clones, along with various amount of α-actin DNA (0.5, 1, 2, or 4 μg) were blotted on Hybond-N+ nylon transfer membrane using the slot-blot apparatus. After denaturation, neutralization and DNA fixation, the filters containing the spotted DNA were hybridized to 32P-labelled cDNA probes. (A) Autoradiogram obtained after Reverse Northern blot hybridization of the spotted cloned gene fragments to a probe prepared by reverse transcription of 30 μg of total RNA purified from the pooled tumour biopsies of six cervical cancer patients. (B) Autoradiogram obtained after Reverse Northern blot hybridization of the spotted cloned gene fragments to a probe prepared by reverse transcription of 30 μg of total RNA purified from the corresponding pooled normal tissue biopsies of the same six cervical cancer employed in (A). (C) The identity and orientation of the cloned gene fragments as spotted on the filter membranes.
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fig2: Reverse Northern blot. DNA (2 μg) of the amplified gene fragment of each of the purified clones, along with various amount of α-actin DNA (0.5, 1, 2, or 4 μg) were blotted on Hybond-N+ nylon transfer membrane using the slot-blot apparatus. After denaturation, neutralization and DNA fixation, the filters containing the spotted DNA were hybridized to 32P-labelled cDNA probes. (A) Autoradiogram obtained after Reverse Northern blot hybridization of the spotted cloned gene fragments to a probe prepared by reverse transcription of 30 μg of total RNA purified from the pooled tumour biopsies of six cervical cancer patients. (B) Autoradiogram obtained after Reverse Northern blot hybridization of the spotted cloned gene fragments to a probe prepared by reverse transcription of 30 μg of total RNA purified from the corresponding pooled normal tissue biopsies of the same six cervical cancer employed in (A). (C) The identity and orientation of the cloned gene fragments as spotted on the filter membranes.

Mentions: 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.


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)

Reverse Northern blot. DNA (2 μg) of the amplified gene fragment of each of the purified clones, along with various amount of α-actin DNA (0.5, 1, 2, or 4 μg) were blotted on Hybond-N+ nylon transfer membrane using the slot-blot apparatus. After denaturation, neutralization and DNA fixation, the filters containing the spotted DNA were hybridized to 32P-labelled cDNA probes. (A) Autoradiogram obtained after Reverse Northern blot hybridization of the spotted cloned gene fragments to a probe prepared by reverse transcription of 30 μg of total RNA purified from the pooled tumour biopsies of six cervical cancer patients. (B) Autoradiogram obtained after Reverse Northern blot hybridization of the spotted cloned gene fragments to a probe prepared by reverse transcription of 30 μg of total RNA purified from the corresponding pooled normal tissue biopsies of the same six cervical cancer employed in (A). (C) The identity and orientation of the cloned gene fragments as spotted on the filter membranes.
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Related In: Results  -  Collection

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getmorefigures.php?uid=PMC2375172&req=5

fig2: Reverse Northern blot. DNA (2 μg) of the amplified gene fragment of each of the purified clones, along with various amount of α-actin DNA (0.5, 1, 2, or 4 μg) were blotted on Hybond-N+ nylon transfer membrane using the slot-blot apparatus. After denaturation, neutralization and DNA fixation, the filters containing the spotted DNA were hybridized to 32P-labelled cDNA probes. (A) Autoradiogram obtained after Reverse Northern blot hybridization of the spotted cloned gene fragments to a probe prepared by reverse transcription of 30 μg of total RNA purified from the pooled tumour biopsies of six cervical cancer patients. (B) Autoradiogram obtained after Reverse Northern blot hybridization of the spotted cloned gene fragments to a probe prepared by reverse transcription of 30 μg of total RNA purified from the corresponding pooled normal tissue biopsies of the same six cervical cancer employed in (A). (C) The identity and orientation of the cloned gene fragments as spotted on the filter membranes.
Mentions: 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.

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