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IL-10 blocks collagen IV invasion by "invasion stimulating factor" activated PC-3 ML cells: upregulation of TIMP-1
M E Stearns1, M Wang, M Stearns
1Medical Colleges of Pennsylvania, Department of Pathology, Philadelphia 19129, USA.
Abstract:
We have previously shown that a 78-kDa "invasion stimulating factor" (ISF) triggers collagenase IV (MMP-2) secretion and the invasive behavior of metastatic PC-3 ML subclones in modified Boyden chamber assays [Stearns, M. E.; Stearns, M. Autocrine factors, type IV collagenase secretion and prostatic cancer cell invasion. Cancer Metastasis Rev. 12:39-52; 1993. Wang, M.; Stearns, M.; Stearns, M. E. Identification of the receptor for a novel M(r) 78,000 "invasion stimulating factor" from metastatic human prostatic PC-3 ML clones. Cancer Res. 54:2492-2495; 1994.]. Recently, we have shown that interleukin 10 (IL-10) preferentially stimulates tissue inhibitor of metalloproteinase-1 (TIMP-1) production in these cells [Wang, M.; Stearns, M. E. Characterization of a novel TIMP-1 enhancer element. J. Biol. Chem., submitted.]. In this paper, we report that IL-10 (20-40 ng) can inhibit the invasion stimulatory effects of ISF (30-60 ng) on PC-3 ML cells. "Checkerboard analysis" with modified Boyden chambers (precoated with 10 and 100 micrograms collagen IV) shows that IL-10 inhibits the stimulatory effects of ISF on both cell motility and chemoinvasion processes. In support of these data, exogenously supplied TIMP-1 (10 micrograms/ml) and collagenase antibodies (1:200 dilution) both completely blocked invasion. Quantitative ELISAs comparing the molar ratios of TIMP-1:MMP-2 and TIMP-2:MMP-2 further demonstrate that IL-10 (10-40 ng) preferentially activates TIMP-1 secretion to increase the molar ratio of TIMP-1:MMP-2 in the presence of increasing amounts of ISF (0-60 ng). IL-10 did not elevate TIMP-2 secretion or influence the molar ratio of TIMP-2:MMP-2.(ABSTRACT TRUNCATED AT 250 WORDS)
Insights
Interleukin 10 (IL-10) inhibits prostate cancer cell invasion by increasing tissue inhibitor of metalloproteinase-1 (TIMP-1) and blocking collagenase IV (MMP-2) activity. This finding offers new therapeutic targets for metastatic prostate cancer.
Area of Science:
- Oncology
- Cell Biology
- Biochemistry
Background:
- Prostate cancer cell invasion is mediated by factors like invasion stimulating factor (ISF) and collagenase IV (MMP-2).
- Interleukin 10 (IL-10) has been shown to stimulate tissue inhibitor of metalloproteinase-1 (TIMP-1) production in these cells.
Purpose of the Study:
- To investigate the inhibitory effects of IL-10 on ISF-induced prostate cancer cell invasion.
- To elucidate the role of TIMP-1 and TIMP-2 in the modulation of invasion by IL-10 and ISF.
Main Methods:
- Modified Boyden chamber assays with collagen IV precoated plates were used for checkerboard analysis.
- Quantitative ELISAs were employed to determine the molar ratios of TIMP-1:MMP-2 and TIMP-2:MMP-2.
- The effects of exogenous TIMP-1 and collagenase antibodies on invasion were assessed.
Main Results:
- IL-10 (20-40 ng) significantly inhibited the invasion-stimulating effects of ISF (30-60 ng) on PC-3 ML cells.
- IL-10 preferentially increased TIMP-1 secretion, elevating the TIMP-1:MMP-2 molar ratio, without affecting TIMP-2 secretion.
- Exogenous TIMP-1 and collagenase antibodies completely blocked cancer cell invasion.
Conclusions:
- IL-10 acts as an inhibitor of prostate cancer cell invasion by modulating the balance between MMP-2 and TIMP-1.
- The findings highlight the potential of targeting the IL-10/TIMP-1 pathway for therapeutic intervention in metastatic prostate cancer.