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Sublethal dose of irradiation enhances invasion of malignant glioma cells through p53-MMP 2 pathway in U87MG mouse brain tumor model.

Pei J, Park IH, Ryu HH, Li SY, Li CH, Lim SH, Wen M, Jang WY, Jung S - Radiat Oncol (2015)

Bottom Line: TIMP-2 expression did not increase in U87MG cells.MMP-2 activity decreased in p53 knock-downed U87MG cells but increased in the control group.Tumor cells invaded by radiation overexpressed MMP-2 and p53 and revealed high gelatinolytic activity compared with those of non-radiated tumor cells.

View Article: PubMed Central - PubMed

Affiliation: Department of Neurosurgery, Chonnam National University Hwasun Hospital and Mediacal School, 322 Seoyang-ro, Hwasun-eup, Hwasun-gun, Jeonnam, 519-763, Republic of Korea.

ABSTRACT

Background: Glioblastoma is a highly lethal neoplasm that frequently recurs locally after radiotherapy, and most of these recurrences originate from near the irradiated target field. In the present study, we identified the effects of radiation on glioma invasion and p53, TIMP-2, and MMP-2 expression through in vitro and in vivo experiments.

Methods: The U87MG (wt p53) and U251 (mt p53) human malignant glioma cell lines were prepared, and the U2OS (wt 53) and Saos2 (del p53) osteosarcoma cell lines were used as p53 positive and negative controls. The four cell lines and p53 knock-downed U87MG cells received radiation (2-6 Gy) and were analyzed for expression of p53 and TIMP-2 by Western blot, and MMP-2 activity was detected by zymography. In addition, the effects of irradiation on directional invasion of malignant glioma were evaluated by implanting nude mice with bioluminescent u87-Fluc in vivo followed by MMP-2, p53, and TIMP-2 immunohisto-chemistry and in situ zymography.

Results: MMP-2 activity and p53 expression increased in proportional to the radiation dose in cell lines with wt p53, but not in the cell lines with del or mt p53. TIMP-2 expression did not increase in U87MG cells. MMP-2 activity decreased in p53 knock-downed U87MG cells but increased in the control group. Furthermore, radiation enhanced MMP-2 activity and increased tumor margin invasiveness in vivo. Tumor cells invaded by radiation overexpressed MMP-2 and p53 and revealed high gelatinolytic activity compared with those of non-radiated tumor cells.

Conclusion: Radiation-induced upregulation of p53 modulated MMP-2 activity, and the imbalance between MMP-2 and TIMP-2 may have an important role in glioblastoma invasion by degrading the extracellular matrix. Bioluminescent "U87-Fluc"was useful for observing tumor formation without sacrifice after implanting tumor cells in the mouse brain. These findings suggest that the radiotherapy involved field for malignant glioma needs to be reconsidered, and that future trials should investigate concurrent pharmacologic therapies that inhibit invasion associated with radiotherapy.

No MeSH data available.


Related in: MedlinePlus

Radiation effect on p53, TIMP2 expression and MMP-2 activity in 2 cell lines with wt p53 and 2 cell lines without p53. Each cell lines were exposed 0, 2, 4, 6 Gy, respectively. MMP-2 activity was detected using zymography (one lane) and expression of TIMP-2 and p53 was detected using western blot (2–4 lane) a Glioblastoma cell lines; b MMP-2 activity of U87MG and U251; c p53 density of U87MG and U251; d Osteosarcoma cell lines
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Fig1: Radiation effect on p53, TIMP2 expression and MMP-2 activity in 2 cell lines with wt p53 and 2 cell lines without p53. Each cell lines were exposed 0, 2, 4, 6 Gy, respectively. MMP-2 activity was detected using zymography (one lane) and expression of TIMP-2 and p53 was detected using western blot (2–4 lane) a Glioblastoma cell lines; b MMP-2 activity of U87MG and U251; c p53 density of U87MG and U251; d Osteosarcoma cell lines

Mentions: The human malignant glioma cell lines U87MG (wt p53) and U251 (mt p53) and the human osteosarcoma cell lines U2OS (wt p53) and SAOS (del p53) were selected to measure the effect of radiation on p53 expression and MMP-2 activity. As shown Fig. 1, MMP-2 activity and p53 expression increased in U87MG and U2OS cells wt p53 in proportion to the radiation dose, but these were unchanged in U251 and SAO2 cells. TIMP-2 is an endogenous MMP-2 inhibitor, and the balance between TIMP-2 and MMP-2 is a critical determinant of malignant glioma invasion. We performed a Western blot for TIMP-2 in the four cell lines exposed to radiation. TIMP-2 expression did not change after radiation exposure in three of the cell lines, except U2OS. The sublethal radiation dose increased p53 expression and MMP-2 activity in U87MG cells without a corresponding increase in TIMP-2 (p < 0.05, Fig. 1a, b, c). In contrast, TIMP-2 expression and MMP-2 activity in U2OS cells with wt p53 increased simultaneously following radiation. These results suggest that TIMP-2 expression was likely to be different depending on the cell type, and that the imbalance between TIMP-2 and MMP-2 expression may enhanced tumor invasion. In addition, MMP-2 activity caused by radiation may be associated with p53 status.Fig. 1


Sublethal dose of irradiation enhances invasion of malignant glioma cells through p53-MMP 2 pathway in U87MG mouse brain tumor model.

Pei J, Park IH, Ryu HH, Li SY, Li CH, Lim SH, Wen M, Jang WY, Jung S - Radiat Oncol (2015)

Radiation effect on p53, TIMP2 expression and MMP-2 activity in 2 cell lines with wt p53 and 2 cell lines without p53. Each cell lines were exposed 0, 2, 4, 6 Gy, respectively. MMP-2 activity was detected using zymography (one lane) and expression of TIMP-2 and p53 was detected using western blot (2–4 lane) a Glioblastoma cell lines; b MMP-2 activity of U87MG and U251; c p53 density of U87MG and U251; d Osteosarcoma cell lines
© Copyright Policy - OpenAccess
Related In: Results  -  Collection

License 1 - License 2
Show All Figures
getmorefigures.php?uid=PMC4554349&req=5

Fig1: Radiation effect on p53, TIMP2 expression and MMP-2 activity in 2 cell lines with wt p53 and 2 cell lines without p53. Each cell lines were exposed 0, 2, 4, 6 Gy, respectively. MMP-2 activity was detected using zymography (one lane) and expression of TIMP-2 and p53 was detected using western blot (2–4 lane) a Glioblastoma cell lines; b MMP-2 activity of U87MG and U251; c p53 density of U87MG and U251; d Osteosarcoma cell lines
Mentions: The human malignant glioma cell lines U87MG (wt p53) and U251 (mt p53) and the human osteosarcoma cell lines U2OS (wt p53) and SAOS (del p53) were selected to measure the effect of radiation on p53 expression and MMP-2 activity. As shown Fig. 1, MMP-2 activity and p53 expression increased in U87MG and U2OS cells wt p53 in proportion to the radiation dose, but these were unchanged in U251 and SAO2 cells. TIMP-2 is an endogenous MMP-2 inhibitor, and the balance between TIMP-2 and MMP-2 is a critical determinant of malignant glioma invasion. We performed a Western blot for TIMP-2 in the four cell lines exposed to radiation. TIMP-2 expression did not change after radiation exposure in three of the cell lines, except U2OS. The sublethal radiation dose increased p53 expression and MMP-2 activity in U87MG cells without a corresponding increase in TIMP-2 (p < 0.05, Fig. 1a, b, c). In contrast, TIMP-2 expression and MMP-2 activity in U2OS cells with wt p53 increased simultaneously following radiation. These results suggest that TIMP-2 expression was likely to be different depending on the cell type, and that the imbalance between TIMP-2 and MMP-2 expression may enhanced tumor invasion. In addition, MMP-2 activity caused by radiation may be associated with p53 status.Fig. 1

Bottom Line: TIMP-2 expression did not increase in U87MG cells.MMP-2 activity decreased in p53 knock-downed U87MG cells but increased in the control group.Tumor cells invaded by radiation overexpressed MMP-2 and p53 and revealed high gelatinolytic activity compared with those of non-radiated tumor cells.

View Article: PubMed Central - PubMed

Affiliation: Department of Neurosurgery, Chonnam National University Hwasun Hospital and Mediacal School, 322 Seoyang-ro, Hwasun-eup, Hwasun-gun, Jeonnam, 519-763, Republic of Korea.

ABSTRACT

Background: Glioblastoma is a highly lethal neoplasm that frequently recurs locally after radiotherapy, and most of these recurrences originate from near the irradiated target field. In the present study, we identified the effects of radiation on glioma invasion and p53, TIMP-2, and MMP-2 expression through in vitro and in vivo experiments.

Methods: The U87MG (wt p53) and U251 (mt p53) human malignant glioma cell lines were prepared, and the U2OS (wt 53) and Saos2 (del p53) osteosarcoma cell lines were used as p53 positive and negative controls. The four cell lines and p53 knock-downed U87MG cells received radiation (2-6 Gy) and were analyzed for expression of p53 and TIMP-2 by Western blot, and MMP-2 activity was detected by zymography. In addition, the effects of irradiation on directional invasion of malignant glioma were evaluated by implanting nude mice with bioluminescent u87-Fluc in vivo followed by MMP-2, p53, and TIMP-2 immunohisto-chemistry and in situ zymography.

Results: MMP-2 activity and p53 expression increased in proportional to the radiation dose in cell lines with wt p53, but not in the cell lines with del or mt p53. TIMP-2 expression did not increase in U87MG cells. MMP-2 activity decreased in p53 knock-downed U87MG cells but increased in the control group. Furthermore, radiation enhanced MMP-2 activity and increased tumor margin invasiveness in vivo. Tumor cells invaded by radiation overexpressed MMP-2 and p53 and revealed high gelatinolytic activity compared with those of non-radiated tumor cells.

Conclusion: Radiation-induced upregulation of p53 modulated MMP-2 activity, and the imbalance between MMP-2 and TIMP-2 may have an important role in glioblastoma invasion by degrading the extracellular matrix. Bioluminescent "U87-Fluc"was useful for observing tumor formation without sacrifice after implanting tumor cells in the mouse brain. These findings suggest that the radiotherapy involved field for malignant glioma needs to be reconsidered, and that future trials should investigate concurrent pharmacologic therapies that inhibit invasion associated with radiotherapy.

No MeSH data available.


Related in: MedlinePlus