Construction of a stable target interfering long non-coding RNA LINC01224 colorectal cancer cell line and its effect on apoptosis
10.19405/j.cnki.issn1000-1492.2022.06.012
- Author:
Guodan Li
1
;
Qin Yang
1
;
Qiuyan Li
1
;
Xiaoyi Chen
1
;
Weiyu Fu
1
;
Liubo Lan
1
;
Xiaocong Lin
1
Author Information
1. Institute of Biochemistry and Molecular Biology, Guangdong Medical University, Zhanjiang 524023
- Publication Type:Journal Article
- Keywords:
colorectal cancer:long non-coding RNA;
lentivirus;
apoptosis
- From:
Acta Universitatis Medicinalis Anhui
2022;57(6):908-914
- CountryChina
- Language:Chinese
-
Abstract:
Objective:To establish colorectal cancer LoVo and SW620 cell lines that stably interfere with the expression of LINC01224, and to explore the effect of down-regulating the expression of LINC01224 on cell apoptosis.
Methods:The GEPIA2 database was used to analyze the expression of LINC01224 in colorectal cancer tissues; qPCR method was used to detect the expression of LINC01224 in 10 human colorectal cancer cells. Three different LINC01224 siRNAs were respectively transfected into human colorectal cancer LoVo cells, and the LINC01224 shRNA lentiviral vector was constructed with the siRNA sequence with the most obvious inhibitory effect of LINC01224 expression. Recombinant lentiviral particles were packaged in HEK293 T cells and then infected with LoVo and SW620 cells. After selection by puromycin, the monoclonal cells that stably interfere with LINC01224 were obtained by limiting dilution method. MTS method detects cell proliferation ability, and flow cytometry detects cell apoptosis rate.
Results:The expression of LINC01224 in colorectal cancer tissues was higher than that in normal colorectal tissues, and its expression in 10 types of colorectal cancer cells was also higher than that in normal colorectal epithelial cells HCOEPic. The inhibition rate of siRNA-3 on the expression of LINC01224 in LoVo cells was higher than that of siRNA-1 and siRNA-2. Therefore, siRNA-3 was chosen to design LINC01224 shRNA.Compared with the control group(sh-NC group), the expression level of LINC01224 in the LoVo and SW620 cells of the stable interference LINC01224 group(sh-LINC01224 group) was reduced(P<0.01), and the cell growth rate was slowed down(P<0.01), the rate of apoptosis also increased(P<0.01).
Conclusion:The shRNA lentiviral interference vector of LINC01224 is successfully constructed, which can stably infect LoVo and SW620 cells, down-regulate the expression of LINC01224 and induce cell apoptosis.
- Full text:2025022111080755528稳定干扰长链非编码RNA_...的建立及其对细胞凋亡的影响_李国丹.pdf
