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Expression and inactivation of glycogen synthase kinase 3 alpha/ beta and their association with the expression of cyclin D1 and p53 in oral squamous cell carcinoma progression.

Mishra R, Nagini S, Rana A - Mol. Cancer (2015)

Bottom Line: The increased expression of pS(21)GSK3α and pS(9)GSK3β not only correlated positively with cyclin D1 and p53 expression in tongue cancer progression but a gradual shift of their expression from the cytoplasmic to the nuclear compartment and overall disease severity was also observed.The interaction of GSK3β-cyclin D1 and the positive correlation of pS(9)GSK3β and the transcription of cyclin D1 were observed.These results demonstrate that the inactivation of GSK3β is an important event in OSCC and can be used as a marker for assessing disease severity and may be exploited for therapeutic intervention.

View Article: PubMed Central - PubMed

Affiliation: Centre for Life Sciences, School of Natural Sciences, Central University of Jharkhand, Ratu-Lohardaga Road, Brambe, Ranchi, 835205, Jharkhand, INDIA. mishrark1@yahoo.co.in.

ABSTRACT

Background: The study aims to evaluate the expression and activity of glycogen synthase kinase 3 isoforms α/β (GSK3α/β) and to assess their oncogenic potential through a correlation with the expression of cyclin D1 and p53 in oral cancer.

Methods: The expression of total and phosphorylated GSK3α/β as well as cyclin D1 and p53 together with their interaction were assessed in human oral cancer tissue samples, apparently normal adjacent tissues, benign tumor samples, premalignant lesions and healthy normal tissues (total 179) using various methods, such as immunohistochemistry, Western blot assays, immunoprecipitation and RT-PCR analysis.

Results: The expression of GSK3β was significantly higher relative to GSK3α indicating the greater role of the β isoform in oral cancer. Among various types of oral cancers, OSCC (of the lip and tongue) showed elevated expression of GSK3α/β, and the expression was correlated with disease progression. The increased expression of pS(21)GSK3α and pS(9)GSK3β not only correlated positively with cyclin D1 and p53 expression in tongue cancer progression but a gradual shift of their expression from the cytoplasmic to the nuclear compartment and overall disease severity was also observed. The interaction of GSK3β-cyclin D1 and the positive correlation of pS(9)GSK3β and the transcription of cyclin D1 were observed.

Conclusions: These results demonstrate that the inactivation of GSK3β is an important event in OSCC and can be used as a marker for assessing disease severity and may be exploited for therapeutic intervention.

No MeSH data available.


Related in: MedlinePlus

The correlation of pSer21GSK3α/pSer9GSK3βexpression with cyclin D1 transcription in various OSCC samples. (A) RT-PCR showing cyclin D1 mRNA expression (201 bp PCR product) in different normals (lane 1–3), PMLs (lane 4–6) and oral cancer samples of various stages (T1-T2 samples lane 7–10; T3-T4 samples lane 11–15). GAPDH expression (410 bp PCR product) was used as a control in this experiment. (B) A histogram showing the level of expression of cyclin D1 mRNA in various groups (N-Normal, PML, T-Tumor) of samples as indicated. No significant correlation (C) of cyclin D1 mRNA expression and pSer21GSK3α protein expression was observed, and a positive correlation (D) between cyclin D1 mRNA expression and the expression of pSer9GSK3β was observed.
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Fig7: The correlation of pSer21GSK3α/pSer9GSK3βexpression with cyclin D1 transcription in various OSCC samples. (A) RT-PCR showing cyclin D1 mRNA expression (201 bp PCR product) in different normals (lane 1–3), PMLs (lane 4–6) and oral cancer samples of various stages (T1-T2 samples lane 7–10; T3-T4 samples lane 11–15). GAPDH expression (410 bp PCR product) was used as a control in this experiment. (B) A histogram showing the level of expression of cyclin D1 mRNA in various groups (N-Normal, PML, T-Tumor) of samples as indicated. No significant correlation (C) of cyclin D1 mRNA expression and pSer21GSK3α protein expression was observed, and a positive correlation (D) between cyclin D1 mRNA expression and the expression of pSer9GSK3β was observed.

Mentions: RT-PCR analysis was performed to determine the expression of cyclin D1 mRNA in various tumor, PML and normal samples (Figure 7A). PMLs and tumor samples showed increased cyclin D1 mRNA levels compared to normal oral mucosa (Figure 7B). The mRNA expression was correlated with the protein expression in the same tissue samples. The correlation of pS21GSK3α expression with cyclin D1 mRNA expression was not significant (n = 28, Pearson’s r = 0.2896, p = 0.135) whereas the correlation of pS9GSK3β expression with cyclin D1 mRNA expression was significant (n = 28, Pearson’s r = 0.8624, p < 0.0001; Figure 7C and D).Figure 7


Expression and inactivation of glycogen synthase kinase 3 alpha/ beta and their association with the expression of cyclin D1 and p53 in oral squamous cell carcinoma progression.

Mishra R, Nagini S, Rana A - Mol. Cancer (2015)

The correlation of pSer21GSK3α/pSer9GSK3βexpression with cyclin D1 transcription in various OSCC samples. (A) RT-PCR showing cyclin D1 mRNA expression (201 bp PCR product) in different normals (lane 1–3), PMLs (lane 4–6) and oral cancer samples of various stages (T1-T2 samples lane 7–10; T3-T4 samples lane 11–15). GAPDH expression (410 bp PCR product) was used as a control in this experiment. (B) A histogram showing the level of expression of cyclin D1 mRNA in various groups (N-Normal, PML, T-Tumor) of samples as indicated. No significant correlation (C) of cyclin D1 mRNA expression and pSer21GSK3α protein expression was observed, and a positive correlation (D) between cyclin D1 mRNA expression and the expression of pSer9GSK3β was observed.
© Copyright Policy - open-access
Related In: Results  -  Collection

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Show All Figures
getmorefigures.php?uid=PMC4334357&req=5

Fig7: The correlation of pSer21GSK3α/pSer9GSK3βexpression with cyclin D1 transcription in various OSCC samples. (A) RT-PCR showing cyclin D1 mRNA expression (201 bp PCR product) in different normals (lane 1–3), PMLs (lane 4–6) and oral cancer samples of various stages (T1-T2 samples lane 7–10; T3-T4 samples lane 11–15). GAPDH expression (410 bp PCR product) was used as a control in this experiment. (B) A histogram showing the level of expression of cyclin D1 mRNA in various groups (N-Normal, PML, T-Tumor) of samples as indicated. No significant correlation (C) of cyclin D1 mRNA expression and pSer21GSK3α protein expression was observed, and a positive correlation (D) between cyclin D1 mRNA expression and the expression of pSer9GSK3β was observed.
Mentions: RT-PCR analysis was performed to determine the expression of cyclin D1 mRNA in various tumor, PML and normal samples (Figure 7A). PMLs and tumor samples showed increased cyclin D1 mRNA levels compared to normal oral mucosa (Figure 7B). The mRNA expression was correlated with the protein expression in the same tissue samples. The correlation of pS21GSK3α expression with cyclin D1 mRNA expression was not significant (n = 28, Pearson’s r = 0.2896, p = 0.135) whereas the correlation of pS9GSK3β expression with cyclin D1 mRNA expression was significant (n = 28, Pearson’s r = 0.8624, p < 0.0001; Figure 7C and D).Figure 7

Bottom Line: The increased expression of pS(21)GSK3α and pS(9)GSK3β not only correlated positively with cyclin D1 and p53 expression in tongue cancer progression but a gradual shift of their expression from the cytoplasmic to the nuclear compartment and overall disease severity was also observed.The interaction of GSK3β-cyclin D1 and the positive correlation of pS(9)GSK3β and the transcription of cyclin D1 were observed.These results demonstrate that the inactivation of GSK3β is an important event in OSCC and can be used as a marker for assessing disease severity and may be exploited for therapeutic intervention.

View Article: PubMed Central - PubMed

Affiliation: Centre for Life Sciences, School of Natural Sciences, Central University of Jharkhand, Ratu-Lohardaga Road, Brambe, Ranchi, 835205, Jharkhand, INDIA. mishrark1@yahoo.co.in.

ABSTRACT

Background: The study aims to evaluate the expression and activity of glycogen synthase kinase 3 isoforms α/β (GSK3α/β) and to assess their oncogenic potential through a correlation with the expression of cyclin D1 and p53 in oral cancer.

Methods: The expression of total and phosphorylated GSK3α/β as well as cyclin D1 and p53 together with their interaction were assessed in human oral cancer tissue samples, apparently normal adjacent tissues, benign tumor samples, premalignant lesions and healthy normal tissues (total 179) using various methods, such as immunohistochemistry, Western blot assays, immunoprecipitation and RT-PCR analysis.

Results: The expression of GSK3β was significantly higher relative to GSK3α indicating the greater role of the β isoform in oral cancer. Among various types of oral cancers, OSCC (of the lip and tongue) showed elevated expression of GSK3α/β, and the expression was correlated with disease progression. The increased expression of pS(21)GSK3α and pS(9)GSK3β not only correlated positively with cyclin D1 and p53 expression in tongue cancer progression but a gradual shift of their expression from the cytoplasmic to the nuclear compartment and overall disease severity was also observed. The interaction of GSK3β-cyclin D1 and the positive correlation of pS(9)GSK3β and the transcription of cyclin D1 were observed.

Conclusions: These results demonstrate that the inactivation of GSK3β is an important event in OSCC and can be used as a marker for assessing disease severity and may be exploited for therapeutic intervention.

No MeSH data available.


Related in: MedlinePlus