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Apolipoprotein M increases the expression of vitamin D receptor mRNA in colorectal cancer cells detected with duplex fluorescence reverse transcription-quantitative polymerase chain reaction

  • Authors:
    • Miao‑Mei Yu
    • Shuang Yao
    • Kai‑Ming Luo
    • Qin‑Feng Mu
    • Yang Yu
    • Guang‑Hua Luo
    • Ning Xu
  • View Affiliations / Copyright

    Affiliations: Comprehensive Laboratory, The Third Affiliated Hospital of Soochow University, Changzhou, Jiangsu 213003, P.R. China, Section of Clinical Chemistry and Pharmacology, Institute of Laboratory Medicine, Lunds University, S‑22185 Lund, Sweden
    Copyright: © Yu et al. This is an open access article distributed under the terms of Creative Commons Attribution License.
  • Pages: 1167-1172
    |
    Published online on: June 7, 2017
       https://doi.org/10.3892/mmr.2017.6716
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Abstract

Apolipoprotein M (ApoM) and the vitamin D receptor (VDR) are apolipoproteins predominantly presenting in high-density lipoprotein (HDL) and a karyophilic protein belonging to the steroid‑thyroid receptor superfamily, respectively. Previous studies have demonstrated that ApoM and VDR are associated with cholesterol metabolism, immune and colorectal cancer regulation. In order to investigate whether ApoM affected the expression of VDR in colorectal cancer cells, a single‑tube duplex fluorescence reverse transcription‑quantitative polymerase chain reaction (RT‑qPCR) system was developed to simultaneously detect the mRNA levels of VDR and GAPDH in HT‑29 cells overexpressing ApoM. The results demonstrated that the amplification products were confirmed as the specific fragment of VDR/GAPDH using the DNA sequencing instrument. The sensitivity, linear range, correlation coefficient, amplification efficiency, intra‑assay and inter‑assay coefficients of variation were 40 copies/µl, 4.00x101‑4.00x105 copies/µl, 0.999, 92.42%, 0.09‑0.34% and 0.32‑0.65% for VDR, and 40 copies/µl, 4.00x101‑4.00x105 copies/µl, 0.999, 98.07%, 0.19‑0.43% and 0.40‑0.75% for GAPDH, respectively. The results indicated that the expression of VDR mRNA was significantly higher in HT‑29 cells overexpressing ApoM, compared with the negative control group (P<0.05). In conclusion, the current study successfully developed the single‑tube duplex RT‑qPCR to simultaneously detect VDR and GAPDH expression in colorectal cancer cells. The methodology results demonstrated that the duplex RT‑qPCR system with high sensitivity and specificity could ensure the objectivity and credibility of the detection. The present study confirmed that ApoM significantly increased the expression of VDR in HT‑29 cells. In addition, it was hypothesized that ApoM may be involved in antineoplastic activity via the upregulation of VDR expression, which may provide novel directions for the investigation of ApoM in cancer.
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Copy and paste a formatted citation
Spandidos Publications style
Yu MM, Yao S, Luo KM, Mu QF, Yu Y, Luo GH and Xu N: Apolipoprotein M increases the expression of vitamin D receptor mRNA in colorectal cancer cells detected with duplex fluorescence reverse transcription-quantitative polymerase chain reaction. Mol Med Rep 16: 1167-1172, 2017.
APA
Yu, M., Yao, S., Luo, K., Mu, Q., Yu, Y., Luo, G., & Xu, N. (2017). Apolipoprotein M increases the expression of vitamin D receptor mRNA in colorectal cancer cells detected with duplex fluorescence reverse transcription-quantitative polymerase chain reaction. Molecular Medicine Reports, 16, 1167-1172. https://doi.org/10.3892/mmr.2017.6716
MLA
Yu, M., Yao, S., Luo, K., Mu, Q., Yu, Y., Luo, G., Xu, N."Apolipoprotein M increases the expression of vitamin D receptor mRNA in colorectal cancer cells detected with duplex fluorescence reverse transcription-quantitative polymerase chain reaction". Molecular Medicine Reports 16.2 (2017): 1167-1172.
Chicago
Yu, M., Yao, S., Luo, K., Mu, Q., Yu, Y., Luo, G., Xu, N."Apolipoprotein M increases the expression of vitamin D receptor mRNA in colorectal cancer cells detected with duplex fluorescence reverse transcription-quantitative polymerase chain reaction". Molecular Medicine Reports 16, no. 2 (2017): 1167-1172. https://doi.org/10.3892/mmr.2017.6716
Copy and paste a formatted citation
x
Spandidos Publications style
Yu MM, Yao S, Luo KM, Mu QF, Yu Y, Luo GH and Xu N: Apolipoprotein M increases the expression of vitamin D receptor mRNA in colorectal cancer cells detected with duplex fluorescence reverse transcription-quantitative polymerase chain reaction. Mol Med Rep 16: 1167-1172, 2017.
APA
Yu, M., Yao, S., Luo, K., Mu, Q., Yu, Y., Luo, G., & Xu, N. (2017). Apolipoprotein M increases the expression of vitamin D receptor mRNA in colorectal cancer cells detected with duplex fluorescence reverse transcription-quantitative polymerase chain reaction. Molecular Medicine Reports, 16, 1167-1172. https://doi.org/10.3892/mmr.2017.6716
MLA
Yu, M., Yao, S., Luo, K., Mu, Q., Yu, Y., Luo, G., Xu, N."Apolipoprotein M increases the expression of vitamin D receptor mRNA in colorectal cancer cells detected with duplex fluorescence reverse transcription-quantitative polymerase chain reaction". Molecular Medicine Reports 16.2 (2017): 1167-1172.
Chicago
Yu, M., Yao, S., Luo, K., Mu, Q., Yu, Y., Luo, G., Xu, N."Apolipoprotein M increases the expression of vitamin D receptor mRNA in colorectal cancer cells detected with duplex fluorescence reverse transcription-quantitative polymerase chain reaction". Molecular Medicine Reports 16, no. 2 (2017): 1167-1172. https://doi.org/10.3892/mmr.2017.6716
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