Teng-Long-Bu-Zhong-Tang induces p21-dependent cell senescence in colorectal carcinoma LS174T cells via histone acetylation

Figures & data

Figure 1 Role of p21 in TLBZT-induced cell senescence.

Notes: After 24 h of transfection with p21 siRNA or control siRNA, LS174T cells were treated with 200 μg/mL of TLBZT for 5 days and subjected to Western blot analyses (A) and cell senescence detection (B). Data are representative of three independent experiments. **P<0.01 vs control, ^#P>0.05 vs TLBZT, ^∆∆P<0.01 vs control siRNA.
Abbreviations: siRNA, small interfering RNA; TLBZT, Teng-Long-Bu-Zhong-Tang.

Figure 2 Role of p53 in TLBZT-induced p21 expression.

Notes: After 24 h of transfection with p53 siRNA or control siRNA, LS174T cells were treated with 200 μg/mL of TLBZT for 5 days and subjected to Western blot probing p53 and p21; β-actin was used as a loading control.
Abbreviations: siRNA, small interfering RNA; TLBZT, Teng-Long-Bu-Zhong-Tang.

Figure 3 Effects of TLBZT on HDAC activity.

Notes: LS174T cells were collected after TLBZT treatment at 100 and 200 μg/mL for 5 days, and subjected to HDAC activity detection according to the manufacturer’s protocol (A). LS174T cells were treated with TLBZT and SVP (200 μM) (B) or TSA (10 nM) (C) for 5 days and subjected to cell senescence detection. Data are from three independent experiments. **P<0.01 vs control, ^∆∆P<0.01 vs SVP (−) or TSA (−).
Abbreviations: HDAC, histone deacetylase; SVP, sodium valproate; TLBZT, Teng-Long-Bu-Zhong-Tang; TSA, trichostatin A.

Figure 4 Effects of TLBZT on H3 and H4 acetylation.

Notes: LS174T cells were treated with 100 and 200 μg/mL of TLBZT for 5 days and subjected to Western blots using indicated antibodies (A). Histone acetylation was quantified by the Quantity One software and expressed as fold of histone (B). **P<0.01 vs control.
Abbreviations: AcH3, acetylated histone H3; AcH4, acetylated histone H4; H3, histone H3; H4, histone H4; TLBZT, Teng-Long-Bu-Zhong-Tang.

Figure 5 Effects of TLBZT on histone H3 and H4 acetylation at the p21 promoter region.

Notes: LS174T cells were treated with 100 and 200 μg/mL of TLBZT for 5 days and subjected to ChIP assay using AcH3 and AcH4 antibodies or control IgG. Precipitated genomic DNA was amplified by qPCR and expressed as a percentage of total input genomic DNA. Data are representative of three independent experiments. **P<0.01 vs control.
Abbreviations: AcH3, acetylated histone H3; AcH4, acetylated histone H4; ChIP, chromatin immunoprecipitation; qPCR, quantitative polymerase chain reaction; TLBZT, Teng-Long-Bu-Zhong-Tang.