RNA sequencing (RNA-seq) of LAIR-1 overexpression in SKOV-3 ovarian cancer cell lines

Leukocyte-associated immunoglobulin-like receptor 1 (LAIR-1), is widely expressed by solid tumor cells and functions as a tumor suppressor. Our previous studies have found that LAIR-1 is expressed in ovarian cancer tumor tissues and in a variety of epithelial ovarian cancer cell lines and that LAIR-1 could inhibit ovarian cancer cell growth by suppressing the PI3K-AKT-mTOR pathway. The aim of the present study was to obtain a comprehensive understanding of the molecular mechanism of LAIR-1 in regulating the biological behavior of ovarian cancer cells. The approach taken was to perform RNA-seq sequencing on SKOV3 cells overexpressing LAIR-1, and to conduct further validation by combinatorial integration of bioinformatics analyses to assess the genetic alterations and using biological experimental findings. Overall design: There are two groups cells used for this expression profile comparison : NC (negative control) and OE(LAIR-1–overexpression) . Total RNA was extracted with Trizol, 1 µg of total RNA was hybridized to the probe and digested with RNase H followed by DNase I. RNA was purified using VAHTS RNA purification beads, 18.5 µL Frag/Prime Buffer (1x) was added, mixed by pipetting, and allowed to stand for 2 minutes at room temperature. After standing on a magnetic stand for 5 minutes to allow the solution to clear, 16 µL of supernatant was gently aspirated into a new nuclease-free centrifuge tube, and the samples were fragmented using a PCR instrument and sequenced using an Illumina sequencer.