GC cell lines (HGC-27, AGS, MKN74, and GTL-16) and the normal human gastric epithelial cell line GES-1 were purchased from ATCC (Manassas, USA). Cells were maintained in RPMI-1640 (HGC-27, MKN74, and GES-1) or Ham’s F-12 (AGS) medium supplemented with 10% fetal bovine serum and antibiotics at 37°C with 5% CO
2.
For transfection, two siRNAs (si-ELFN1-AS1#1 and si-ELFN1-AS1#2), an overexpression vector of ELFN1-AS1 (OE-lnc), miR-211-3p mimic/inhibitor, siRNA-TRIM29, and their negative controls (NC) were purchased from GenePharma (Guangzhou, China). All sequences of siRNAs are listed in
Table 3 . Then, 100 nM of the aforementioned vectors were transfected or cotransfected into AGS and MKN74 cells (1×10
6) using Lipofectamine 3000 (Invitrogen) according to the manufacturer’s instructions. qRT-PCR was performed to assess the transfection efficiency after 48 h.
2.
For transfection, two siRNAs (si-ELFN1-AS1#1 and si-ELFN1-AS1#2), an overexpression vector of ELFN1-AS1 (OE-lnc), miR-211-3p mimic/inhibitor, siRNA-TRIM29, and their negative controls (NC) were purchased from GenePharma (Guangzhou, China). All sequences of siRNAs are listed in
6) using Lipofectamine 3000 (Invitrogen) according to the manufacturer’s instructions. qRT-PCR was performed to assess the transfection efficiency after 48 h.
Name | Sequence |
si-ELFN1-AS1#1 | 5′-GCUUGUGGUUUCUCACAAA-3′ |
si-ELFN1-AS1#2 | 5′-CCUUUAAUCUCUUGCUCAA-3′ |
si-TRIM29 | 5′-GGGACAAGACTATGCTTTA-3′ |
si-NC | 5′-UUCUCCGAACGUGUCACGUTT-3′ |
miR-211-3p mimic | 5′-GCAGGGACAGCAAAGGGGUGC-3′ |
Mimic-NC | 5′-UUCUCCGAACGUGUCACG-3′ |
miR-211-3p inhibitor | 5′-UACAAAAGGGUAGUGAAAGUA-3′ |
Inhibitor-NC | 5′- UAGAACUUGCAUUGCAACCGG-3′ |