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Published on: June 2, 2016
Survivin gene RNA interference induces apoptosis in human HL60 leukemia cell lines
Yun-Hua Lu1, Xue-Gang Luo, Xi Tao
1Research Centre of Biotechnology, China Pharmaceutical University, NanJing, China.
Background:
It has been demonstrated that survivin, a member of the inhibitor of apoptosis (IAP) protein family, is expressed in human cancers but is undetectable in normal differentiated tissues. HL60 siRNA was introduced into HL60 cells to investigate its effect on cancer cell growth.
Methods:
The opposite livability on HL60 cells was assayed with an MTT test. The change of mRNA and protein of the survivin gene were detected by reverse transcriptase polymerase chain reaction and Western blot, respectively. Cell apoptosis was assayed by flow cytometry.
Results:
The growth of HL60 cells decreased by 65.3%, 62.1%, and 52.4% when they transfected with 400 nM siRNA lyh1, lyh2, and lyh3 after 48 hours, in comparison to the control group. Also, the mRNA and protein were knocked down and HL60 cells presented an increase in apoptosis index.
Conclusions:
Small interfering RNA can exert a knockdown of survivin gene expression in HL60 cells, and effectively induce apoptosis and inhibit the growth of leukemia cells.
Insights
Small interfering RNA (siRNA) targeting survivin effectively inhibited leukemia cell growth and induced apoptosis. This study demonstrates siRNA's potential in cancer therapy by reducing survivin expression in HL60 cells.
Area of Science:
- Molecular Biology
- Cancer Research
- Gene Therapy
Background:
- Survivin, an inhibitor of apoptosis protein (IAP) family member, is upregulated in various human cancers.
- Survivin is typically absent in normal differentiated tissues, making it a potential cancer-specific target.
- Investigating survivin's role in cancer cell proliferation is crucial for developing targeted therapies.
Purpose of the Study:
- To investigate the effect of survivin small interfering RNA (siRNA) on the growth of HL60 leukemia cells.
- To determine if survivin gene knockdown influences HL60 cell apoptosis.
- To assess the efficacy of siRNA-mediated survivin inhibition in a leukemia cell model.
Main Methods:
- HL60 cells were transfected with varying concentrations of survivin siRNA (lyh1, lyh2, lyh3).
- Cell viability was assessed using the MTT assay.
- Survivin mRNA and protein levels were quantified via RT-PCR and Western blot, respectively.
- Apoptosis was analyzed using flow cytometry.
Main Results:
- Transfection with 400 nM siRNA resulted in significant HL60 cell growth inhibition (52.4%–65.3%) after 48 hours compared to controls.
- Survivin mRNA and protein expression were effectively reduced (knocked down) by siRNA.
- An increased apoptosis index was observed in HL60 cells treated with survivin siRNA.
Conclusions:
- Small interfering RNA (siRNA) can successfully achieve knockdown of survivin gene expression in leukemia cells.
- Survivin knockdown by siRNA effectively induces apoptosis in HL60 cells.
- siRNA-mediated inhibition of survivin demonstrates potential for inhibiting leukemia cell growth.
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