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Updated: Aug 29, 2026
In Vivo Inhibition of MicroRNA to Decrease Tumor Growth in Mice
Published on: August 23, 2019
[Inhibition of survivin expression in liver cancer cells by shRNA]
Ge Yan1, Ai-long Huang, Ni Tang
1Institute for Viral Hepatitis, Chongqing University of Medical Sciences, Chongqing 400010, China.
Objective:
To construct the plasmid containing short hairpin RNA (shRNA) of survivin in order to suppress the expression of survivin gene in HepG2 and SMMC-7721.
Methods:
Two 20 to 21 bp reverse repeated motifs of survivin target sequence with 4 bp or 8 bp spacer were synthesized respectively and inserted into plasmid pTZU6+1 to generate the plasmid pshRNA-survivin1 and pshRNA-survivin2; plasmid pEGFP-C1-survivin and pshRNA-survivin1 or pshRNA-survivin2 plasmid were cotransfected into liver cancer cell HepG2 and SMMC-7721 to detect effect of GFP expression respectively and analyze the inhibition of survivin gene.
Results:
The recombinant plasmid pshRNA-survivin1 and pshRNA-survivin2 were successfully constructed. The recombinant plasmids suppress the survivin expression by 80% in HepG2 and SMMC-7721.
Conclusion:
The result showed that the short hairpin RNA of survivin can efficiently suppress it's expression in HepG2 and SMMC-7721.
Insights
Researchers developed survivin short hairpin RNA (shRNA) plasmids to suppress survivin gene expression in liver cancer cells. The constructs effectively reduced survivin levels by 80% in HepG2 and SMMC-7721 cells.
Area of Science:
- Molecular Biology
- Cancer Research
- Gene Silencing
Background:
- Survivin is a key protein implicated in cell proliferation and apoptosis resistance in various cancers.
- Targeting survivin expression is a potential therapeutic strategy for liver cancer.
Purpose of the Study:
- To construct survivin-targeting short hairpin RNA (shRNA) plasmids.
- To evaluate the efficacy of these shRNA plasmids in suppressing survivin gene expression in liver cancer cell lines.
Main Methods:
- Synthesis and cloning of survivin target sequences into the pTZU6+1 plasmid to create pshRNA-survivin1 and pshRNA-survivin2.
- Co-transfection of these plasmids with pEGFP-C1-survivin into HepG2 and SMMC-7721 cells.
- Assessment of survivin gene inhibition via GFP expression analysis.
Main Results:
- Successfully constructed recombinant plasmids pshRNA-survivin1 and pshRNA-survivin2.
- Achieved an 80% suppression of survivin expression in both HepG2 and SMMC-7721 cell lines.
Conclusions:
- Survivin-targeting shRNA demonstrates significant potential for gene silencing in liver cancer.
- The developed shRNA constructs efficiently suppress survivin expression in HepG2 and SMMC-7721 cells.
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