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Inducible and Reversible Dominant-negative DN Protein Inhibition
Published on: January 7, 2019
Effect on proliferation and apoptosis of retinoblastoma cell by RNA inhibiting high mobility group protein box-1
Li-Lun Wang1, Yan-Qin Feng1, Yu-Hong Cheng2
1Department of Ophthalmology, Affiliated Hospital of Yan'an University, Yan'an 716000, Shaanxi Province, China.
Aim:
To investigate the effect of high mobility group protein box-1 (HMGB1) siRNA on proliferation and apoptosis of retinoblastoma (Rb) cells.
Methods:
The expression of HMGB1 in Rb cells were detected by real-time polymerase chain reaction (RT-PCR) and Western blot. Chemically synthesized HMGB1 siRNA was transfected into Y79 cells. The inhibitory rate was also examined by RT-PCR and Western blot. After HMGB1 siRNA transfection, the cell proliferation was analyzed by MTT, and cell apoptosis was detected by Caspase-3 active detection kit. Cell cycle distribution and apoptosis were detected by flow cytometry.
Results:
The expression of HMGB1 significantly elevated in Rb cells (P<0.01). After transfected by siRNA, the HMGB1 protein level of Y79 cells was significantly reduced (P<0.01). After siRNA interference HMGB1, the proportion of proliferating cells reduced, and the proportion of quiescent cells increased (P<0.05). In addition, apoptosis rate of Y79 cells increased from 2.03% to 9.10% after interfering with HMGB1 siRNA (P<0.05).
Conclusion:
Specific HMGB1 siRNA can inhibit the expression of HMGB1. The effect may be attributed to inhibit the proliferation and promote cell apoptosis.
Insights
High mobility group protein box-1 (HMGB1) siRNA effectively reduces HMGB1 expression in retinoblastoma cells. This inhibition suppresses cell proliferation and promotes apoptosis, offering a potential therapeutic strategy.
Area of Science:
- Ophthalmology
- Molecular Biology
- Cancer Research
Background:
- Retinoblastoma (Rb) is a common intraocular malignancy in children.
- High mobility group protein box-1 (HMGB1) is implicated in various cancers, including Rb.
Purpose of the Study:
- To investigate the therapeutic potential of HMGB1 siRNA in retinoblastoma.
- To determine the effects of HMGB1 inhibition on Rb cell proliferation and apoptosis.
Main Methods:
- HMGB1 expression was quantified using RT-PCR and Western blot.
- Y79 retinoblastoma cells were transfected with HMGB1 siRNA.
- Cell proliferation, apoptosis, and cell cycle distribution were analyzed via MTT assay, Caspase-3 assay, and flow cytometry.
Main Results:
- HMGB1 expression was significantly elevated in Rb cells.
- HMGB1 siRNA transfection markedly reduced HMGB1 protein levels in Y79 cells.
- siRNA-mediated HMGB1 inhibition led to decreased proliferation and increased apoptosis (from 2.03% to 9.10%) in Y79 cells.
Conclusions:
- Specific HMGB1 siRNA effectively inhibits HMGB1 expression in retinoblastoma cells.
- HMGB1 suppression by siRNA demonstrates anti-proliferative and pro-apoptotic effects.
- Targeting HMGB1 presents a promising therapeutic avenue for retinoblastoma treatment.
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