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Abstract
Background
Transforming growth factor-β1 (TGF-β1) is known to be a potent growth inhibitor of many cell types, including most epithelial cells. However, the mechanism of TGF-β1 action on cell growth in lymphomas and leukemia still remains to be elucidated. c-myc is a central regulator of cell proliferation and apoptosis, and telomerase is believed to play an important role in carcinogenesis. The aim of the study was to determine the effects of cell growth, c-myc gene expression and telomerase activity due to TGF-β1 and examine its mechanism of action in lymphomas and leukemia.
Methods
The cell growths of Jiyoye (Burkitt lymphoma), H9 (T cell lymphoma), and CCRF-CEM (acute lymphocytic leukemia, T cell) cell lines due to TGF-β1 were measured using the MTT assay. RT-PCR was also performed to monitor the expression of the c-myc gene in these cells with the telomerase activity measured using a TRAP assay.
Results
There was significant inhibition of cell growth in TGF-β1 (5ng/mL) treated Jiyoye cells. When treated with TGF-β1, the Jiyoye cells exhibited marked decreases in the levels of c-myc RNA and telomerase activity. However, TGF-β1 treated H9 and CCRF-CEM cells showed no cell growth inhibition or reductions in the levels of c-myc mRNA and telomerase activity. The effect of TGF-β1 on cell growth was noted to closely correlate with c-myc mRNA expression and telomerase activity.
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Fig. 1.
Effect of TGF-β1 treatment on J iyoye cell growth. (A) C ontrol cell after 24 hours, (B) 24 hours after treatment of TGF-β1. (C) Control cell after 48 hours, (D) 48 hours after treatment of TGF-β1. (E) Control cell after 72 hours, (F) 72 hours after treatment of TGF-β1.
Fig. 2.
Effect of TGF-β1 treatment on c-myc RNA expression in J iyoye cell line, H9 cell line, and C C RF-C EM cell line. 1, negative control; 2, control after 24 hours; 3, 24 hours after treatment of TGF-β1; 4, control after 48 hours; 5, 48 hours after treatment of TGF-β1; 6, control after 72 hours; 7, 72 hours after treatment of TGF-β1.
Fig. 3.
Absorbance units (A450-A690) of J iyoye cell line for telomerase activity. TRS8, positive control; CHAPS, negative control; C, control; T, TGF-β1 treated.
Fig. 4.
Absorbance units (A450-A690) of H9 cell line for telomerase activity. TRS8, positive control; CHAPS, negative control; C, control; T, TGF-β1 treated.
Fig. 5.
Absorbance units (A450-A690) of C C RF-C EM cell line for telomerase activity. TRS8, positive control; CHAPS, negative control; C, control; T, TGF-β1 treated.
Table 1.
Effect on cell growth by TGF-β1 in J iyoye, H9 and C CRF-C EM cell lines
| Cell survival ratio (%) | P value | |||
|---|---|---|---|---|
| 24 hours | 48 hours 7 | 72 hours | ||
| Control | 100 | 100 | 100 | |
| J iyoye | 88.5 | 70.5 | 56.4 | ＜0.005 |
| H9 | 73.4 | 85.1 | 93.4 | |
| CC RF-CEM | 72.9 | 100 | 114.1 | |
Table 2.
Telomerase activity in J iyoye, H9 and CC RF-CEM cell lines
| Telomerase activity (ΔA∗) | ||||||
|---|---|---|---|---|---|---|
| 24 hours | 48 hours | 72 hours | ||||
| C ontrol | TGF-β1 | C ontrol | TGF-β1 | C ontrol | TGF-β1 | |
| J iyoye | + (0.499) | + (0.230) | + (0.428) | + (0.211) | + (0.404) | – (0.146) |
| H9 | + (0.399) | + (0.324) | + (0.296) | + (0.272) | + (0.222) | + (0.227) |
| C C RF-C EM | + (0.556) | + (0.360) | + (0.554) | + (0.541) | + (0.467) | + (0.467) |
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