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Galectin-3 and Beclin1/Atg6 genes in human cancers: using cDNA tissue panel, qRT-PCR, and logistic regression model to identify cancer cell biomarkers.

Idikio HA - PLoS ONE (2011)

Bottom Line: Miner software derived formula was used to calculate mRNA levels and then fold changes.High levels of Beclin1 mRNA levels were in liver and prostate cancers when compared to normal tissues.Breast, kidney and thyroid cancers had high galectin-3 levels and low Beclin1 levels.

View Article: PubMed Central - PubMed

Affiliation: Department of Laboratory Medicine and Pathology, University of Alberta, Edmonton, Alberta, Canada. hidikio@ualberta.ca

ABSTRACT

Background: Cancer biomarkers are sought to support cancer diagnosis, predict cancer patient response to treatment and survival. Identifying reliable biomarkers for predicting cancer treatment response needs understanding of all aspects of cancer cell death and survival. Galectin-3 and Beclin1 are involved in two coordinated pathways of programmed cell death, apoptosis and autophagy and are linked to necroptosis/necrosis. The aim of the study was to quantify galectin-3 and Beclin1 mRNA in human cancer tissue cDNA panels and determine their utility as biomarkers of cancer cell survival.

Methods and results: A panel of 96 cDNAs from eight (8) different normal and cancer tissue types were used for quantitative real-time polymerase chain reaction (qRT-PCR) using ABI7900HT. Miner2.0, a web-based 4- and 3-parameter logistic regression software was used to derive individual well polymerase chain reaction efficiencies (E) and cycle threshold (Ct) values. Miner software derived formula was used to calculate mRNA levels and then fold changes. The ratios of cancer to normal tissue levels of galectin-3 and Beclin1 were calculated (using the mean for each tissue type). Relative mRNA expressions for galectin-3 were higher than for Beclin1 in all tissue (normal and cancer) types. In cancer tissues, breast, kidney, thyroid and prostate had the highest galectin-3 mRNA levels compared to normal tissues. High levels of Beclin1 mRNA levels were in liver and prostate cancers when compared to normal tissues. Breast, kidney and thyroid cancers had high galectin-3 levels and low Beclin1 levels.

Conclusion: Galectin-3 expression patterns in normal and cancer tissues support its reported roles in human cancer. Beclin1 expression pattern supports its roles in cancer cell survival and in treatment response. qRT-PCR analysis method used may enable high throughput studies to generate molecular biomarker sets for diagnosis and predicting cancer treatment response.

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Beclin1/Atg6 relative mRNA levels in Human Normal and Cancer Tissues.(A). Box plot of the overall expression in all tissue types. (B) Box plots of Beclin1 distribution of mRNA levels in normal tissues (Na = normal breast, Nb = Normal colon, Nc = Normal kidney, Nd = Normal liver, Ne = Normal lung, Nf = Normal ovary, Ng = Normal Prostate, Nh = Normal thyroid). (C) Box-whisker plots of the relative Beclin1 mRNA levels in cancer tissues. BrCa = Breast cancer, Col_Ca = colon cancer, Ca_Kid = Kidney cancer, Ca_Liv = liver cancer, Ca_Lun = lung cancer, Ca_Ova = ovarian cancer, Ca_Pros = Prostate cancer, Ca_Thyr = thyroid cancer (see Table 3a and b for values).
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pone-0026150-g002: Beclin1/Atg6 relative mRNA levels in Human Normal and Cancer Tissues.(A). Box plot of the overall expression in all tissue types. (B) Box plots of Beclin1 distribution of mRNA levels in normal tissues (Na = normal breast, Nb = Normal colon, Nc = Normal kidney, Nd = Normal liver, Ne = Normal lung, Nf = Normal ovary, Ng = Normal Prostate, Nh = Normal thyroid). (C) Box-whisker plots of the relative Beclin1 mRNA levels in cancer tissues. BrCa = Breast cancer, Col_Ca = colon cancer, Ca_Kid = Kidney cancer, Ca_Liv = liver cancer, Ca_Lun = lung cancer, Ca_Ova = ovarian cancer, Ca_Pros = Prostate cancer, Ca_Thyr = thyroid cancer (see Table 3a and b for values).

Mentions: Relative mRNA levels for Beclin1, in all 96 tissue types were lower compared to galectin-3 (Fig. 2a). Minimum value was 1.27e-05 and maximum value was 5.15e-01 (mean = 3.18056e-02+/−6.4003e-02). The distribution of Beclin1 mRNA in normal tissues is presented in Fig. 2b and for cancer tissue types is shown in Fig. 2c and Tables 4 and 5. Colon, prostate, and liver cancer tissues had high relative mRNA values.


Galectin-3 and Beclin1/Atg6 genes in human cancers: using cDNA tissue panel, qRT-PCR, and logistic regression model to identify cancer cell biomarkers.

Idikio HA - PLoS ONE (2011)

Beclin1/Atg6 relative mRNA levels in Human Normal and Cancer Tissues.(A). Box plot of the overall expression in all tissue types. (B) Box plots of Beclin1 distribution of mRNA levels in normal tissues (Na = normal breast, Nb = Normal colon, Nc = Normal kidney, Nd = Normal liver, Ne = Normal lung, Nf = Normal ovary, Ng = Normal Prostate, Nh = Normal thyroid). (C) Box-whisker plots of the relative Beclin1 mRNA levels in cancer tissues. BrCa = Breast cancer, Col_Ca = colon cancer, Ca_Kid = Kidney cancer, Ca_Liv = liver cancer, Ca_Lun = lung cancer, Ca_Ova = ovarian cancer, Ca_Pros = Prostate cancer, Ca_Thyr = thyroid cancer (see Table 3a and b for values).
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Related In: Results  -  Collection

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pone-0026150-g002: Beclin1/Atg6 relative mRNA levels in Human Normal and Cancer Tissues.(A). Box plot of the overall expression in all tissue types. (B) Box plots of Beclin1 distribution of mRNA levels in normal tissues (Na = normal breast, Nb = Normal colon, Nc = Normal kidney, Nd = Normal liver, Ne = Normal lung, Nf = Normal ovary, Ng = Normal Prostate, Nh = Normal thyroid). (C) Box-whisker plots of the relative Beclin1 mRNA levels in cancer tissues. BrCa = Breast cancer, Col_Ca = colon cancer, Ca_Kid = Kidney cancer, Ca_Liv = liver cancer, Ca_Lun = lung cancer, Ca_Ova = ovarian cancer, Ca_Pros = Prostate cancer, Ca_Thyr = thyroid cancer (see Table 3a and b for values).
Mentions: Relative mRNA levels for Beclin1, in all 96 tissue types were lower compared to galectin-3 (Fig. 2a). Minimum value was 1.27e-05 and maximum value was 5.15e-01 (mean = 3.18056e-02+/−6.4003e-02). The distribution of Beclin1 mRNA in normal tissues is presented in Fig. 2b and for cancer tissue types is shown in Fig. 2c and Tables 4 and 5. Colon, prostate, and liver cancer tissues had high relative mRNA values.

Bottom Line: Miner software derived formula was used to calculate mRNA levels and then fold changes.High levels of Beclin1 mRNA levels were in liver and prostate cancers when compared to normal tissues.Breast, kidney and thyroid cancers had high galectin-3 levels and low Beclin1 levels.

View Article: PubMed Central - PubMed

Affiliation: Department of Laboratory Medicine and Pathology, University of Alberta, Edmonton, Alberta, Canada. hidikio@ualberta.ca

ABSTRACT

Background: Cancer biomarkers are sought to support cancer diagnosis, predict cancer patient response to treatment and survival. Identifying reliable biomarkers for predicting cancer treatment response needs understanding of all aspects of cancer cell death and survival. Galectin-3 and Beclin1 are involved in two coordinated pathways of programmed cell death, apoptosis and autophagy and are linked to necroptosis/necrosis. The aim of the study was to quantify galectin-3 and Beclin1 mRNA in human cancer tissue cDNA panels and determine their utility as biomarkers of cancer cell survival.

Methods and results: A panel of 96 cDNAs from eight (8) different normal and cancer tissue types were used for quantitative real-time polymerase chain reaction (qRT-PCR) using ABI7900HT. Miner2.0, a web-based 4- and 3-parameter logistic regression software was used to derive individual well polymerase chain reaction efficiencies (E) and cycle threshold (Ct) values. Miner software derived formula was used to calculate mRNA levels and then fold changes. The ratios of cancer to normal tissue levels of galectin-3 and Beclin1 were calculated (using the mean for each tissue type). Relative mRNA expressions for galectin-3 were higher than for Beclin1 in all tissue (normal and cancer) types. In cancer tissues, breast, kidney, thyroid and prostate had the highest galectin-3 mRNA levels compared to normal tissues. High levels of Beclin1 mRNA levels were in liver and prostate cancers when compared to normal tissues. Breast, kidney and thyroid cancers had high galectin-3 levels and low Beclin1 levels.

Conclusion: Galectin-3 expression patterns in normal and cancer tissues support its reported roles in human cancer. Beclin1 expression pattern supports its roles in cancer cell survival and in treatment response. qRT-PCR analysis method used may enable high throughput studies to generate molecular biomarker sets for diagnosis and predicting cancer treatment response.

Show MeSH
Related in: MedlinePlus