Long noncoding RNA small nucleolar RNA host gene 1 promotes gemcitabine resistance by targeting microRNA330 in pancreatic cancer cells
10.3760/cma.j.cn115667-20201225-00214
- VernacularTitle:长链非编码RNA小核仁RNA宿主基因1靶向miR-330促进胰腺癌细胞吉西他滨耐药作用
- Author:
Rui LI
1
;
Xiping ZHU
;
Lei SONG
Author Information
1. 黑龙江省医院消化病院消化三科，哈尔滨 150086
- Keywords:
Pancreatic neoplasms;
long noncoding RNA;
Drogresistance, neoplasm;
Apoptosis
- From:
Chinese Journal of Pancreatology
2021;21(5):359-364
- CountryChina
- Language:Chinese
-
Abstract:
Objective:To investigate the role of long noncoding RNA (lncRNA) small nucleolar RNA host gene 1 (SNHG1) in the resistance of pancreatic cancer PANC1 cells to gemcitabine (GEM), and clarify the relationship between SNHG1 and miR-330.Methods:The clinical data of 179 pancreatic tissue samples and 171 adjacent normal pancreas tissues were collected from the Cancer Genome Atlas (TCGA), and bioinformatics analysis was performed to analyze the expression of SNHG1 and miR-330 in pancreatic cancer tissue and adjacent normal tissue. PANC1 cell line with the resistance to GEM was established by the intermittent gradient doubling method in vitro. The GEM-resistant cells were divided into three groups: si-NC-GEM (negative control) group, si-SNHG1-GEM (transfected with siRNA targeting SNHG1) group, and GEM-resistant group. At the same time, to validate the role of miR-330 in targeting SNHG1 expression, the GEM-resistant cells were further divided into three groups: NC-inh+ si-NC group (co-transfected with negative control miR-330 inhibitor and si-NC), NC-inh+ si-SNHG1 group (co-transfected with negative control miR-330 inhibitor and si-SNHG1), and miR-330-inh + si-SNHG1 group (co-transfected with miR-330 inhibitor and si-SNHG1). qRT-PCR was used to detect the expression of SNHG1 and miR-330 in GEM-resistant cells. CCK-8 and immunofluorescence (Ki67) were used to test GEM-resistant cell growth and proliferation. Transwell and wound healing were used to test the migration and invasion in GEM-resistant cells. Bioinformatics analysis and luciferase reporter assay were used to verify the regulatory relationship between SNHG1 and miR-330. Results:Compared to the adjacent normal tissues of pancreatic cancer, the expression of lncRNA SNHG1 in pancreatic cancer tissues was significantly increased (6.543±0.72 vs 5.31±0.96), and the expression of miR-330 was greatly decreased (2.54±1.85 vs 3.01±1.23); and all the differences were statistically significant ( P<0.05). The expression of lncRNA SNHG1 in si-NC-GEM group, si-SNHG1-GEM group, and GEM-resistant group was 2.43±0.10, 0.26±0.08 and 3.25±0.310, which in si-SNHG1-GEM group was lower than those in si-NC-GEM group or GEM-resistant group. While the expression of miR-330 in si-NC-GEM group, si-SNHG1-GEM group, and GEM-resistant group was 0.47±0.13, 0.84±0.12 and 0.38±0.21, which in si-SNHG1-GEM group was higher than those in si-NC-GEM group or GEM-resistant group. All the differences were statistically significant ( P<0.05). Compared to the si-NC-GEM group or GEM-resistant group, A450, Ki67, number of migrated cells and the distance of invasion in si-SNHG1-GEM group were all decreased, and the differences were statistically significant ( P<0.05). Otherwise, luciferase activity of miR-330-WT in si SNHG1-GEM group was significantly higher than that in NC siRNA group (3.21±0.22 vs 1.03±0.18). The luciferase activity of SNHG1-WT in miR-330 inhibitor group was significantly lower than that in NC inhibitor group (0.97±0.21 vs 2.32±0.17). In miR-330-inh+ si-SNHG1 GEM-resistant group, the cell A450, Ki67, migration cell and distant of invasion was higher than those in NC-inh+ si-SNHG1 group, and the difference was statistically significant (all P value<0.05). Conclusions:lncRNA SNHG1 targeting miR-330 could promote GEM resistance in pancreatic cancer PANC1 cells.
