Establishment of quantitative nested-PCR of Abelson interactor 1 transcript variant-11

Tingru Lin; Na Wu; Jingzhu Guo; Mei Li; Zhaohui Zhong; Weidong Yu

doi:10.1016/j.heliyon.2022.e12119

Establishment of quantitative nested-PCR of Abelson interactor 1 transcript variant-11

Heliyon. 2022 Dec 7;8(12):e12119. doi: 10.1016/j.heliyon.2022.e12119. eCollection 2022 Dec.

Authors

Tingru Lin^{1

2}, Na Wu¹, Jingzhu Guo³, Mei Li¹, Zhaohui Zhong⁴, Weidong Yu¹

Affiliations

¹ Department of Central Laboratory and Institute of Clinical Molecular Biology, Peking University People's Hospital, Beijing 100044, China.
² Department of Gastroenterology, Peking University People's Hospital, Beijing 100044, China.
³ Department of Pediatric, Peking University People's Hospital, Beijing 100044, China.
⁴ Department of General Surgery, Peking University People's Hospital, Beijing 100044, China.

Abstract

Abelson interactor 1 (ABI1), which presents 18 Transcript Variants (TSV), plays an important role in CRC metastasis. Different ABI1-TSVs play synergistic or antagonistic roles in the same pathophysiological events. ABI1 Transcript Variant-11 (ABI1-TSV-11) functionally promotes lymph node metastasis of left-sided colorectal cancer (LsCC) and is an independent molecular marker to evaluate the prognosis of patients with LsCC. However, there is still lack of a quick and accurate method to detect the expression of ABI-TSV-11, distinguishing ABI1-TSV-11 from other 17 TSVs. To establish a rapid method specific for ABI1-TSV-11detection, we developed a quantitative nested-PCR method composed of pre-amplification regular PCR using ABI1 universal primer pair and the followed Real Time (RT)-qPCR using ABI1-TSV-11 specific primer pair spanning exon-exon junction. ABI1-TSV-11-overexpressed SW480 and LoVo cell lines were used to verify the quantitative nested-PCR assay, and the sequencing data was used to evaluate the accuracy of ABI1-TSV-11 quantitative nested-PCR assay. The detection limit was 5.24×10⁴ copies/ml. ABI1-TSV-11 quantitative nested-PCR provides a new technical means for the detection of ABI1-TSV-11.

Keywords: ABI1 universal primer pair; ABI1-TSV-11; CRC; Exon-exon junction spanning primer; Quantitative nested polymerase chain reaction (quantitative -nested-PCR).