Anti-tumor and anti-invasion effects of a combination of 4-methylumbelliferone and ionizing radiation in human fibrosarcoma cells

Abstract

Abstract. Hyaluronan (HA) is a major component of the extracellular matrix that is synthesized in excess in cancer tissues. 4-methylumbelliferone (MU) inhibits the synthesis of HA and is closely related to the invasion and metastasis of cancer. However, the effects of MU in conjunction with cancer radiotherapy remain unknown. The present study assessed the anti-tumor and anti-invasion effects of the concomitant use of ionizing radiation (IR) and 100 µM MU on human fibrosarcoma HT1080 cells. Cell viability and cellular invasion potency assays were performed. There was a greater decrease in the viability of cells cultured with a combination of 2 Gy IR and MU compared with untreated control cells. In addition, cell cycle distribution analysis demonstrated that a higher proportion of these cells were in the sub-G1 phase and higher fractions of annexin-V positive, propidium iodide positive cells (i.e., apoptotic cells) were observed. HA concentration in the 2 Gy irradiated culture was similar to that in the non-irradiated control culture, however, it significantly decreased following the administration of both MU alone and 2 Gy IR with MU. Furthermore, treatment with 2 Gy IR and MU resulted in a significant decrease in the invasion rate and matrix metalloproteinase (MMP)-2 and MPP-9 expression. Taken together, these results suggest that the administration of MU with 2 Gy IR is effective at reducing HA production, cell invasion and the metastatic potential of cancer cells.

Extracellular Matrix in the Tumor Microenvironment and Its Impact on Cancer Therapy

“…The authors attributed the improved response to both immunotherapy modalities to an increased Tcell infiltration rate after the hyaluronidase treatment. A positive effect of 4-MU treatment was also reported on the sensitivity of fibrosarcoma cells to radiotherapy (Saga et al, 2017). Under the trade name Hychromone, 4-MU is already marketed in Asia and Europe as a choleretic agent (reviewed in Kudo et al, 2017).…”

4‑methylumbelliferone inhibits clonogenic potency by suppressing high molecular weight‑hyaluronan in fibrosarcoma cells

“…HA density quantitation. HA concentration was measured by using the Hyaluronan Quantikine ® ELISA kit (R&D Systems, Inc.) using the procedure reported previously (15). HA concentration was calculated from a standard curve of the measured at 450 nm absorbance.…”

“…RNA extraction and analysis. Total RNA was extracted from HT1080 cells, and RNA quality was confirmed according to the procedure reported previously (15).…”

“…In our previous study, administration of a combination of 100 µM 4-MU and 2 Gy X-ray irradiation caused downregulation of matrix metalloproteinases-2 and -9 and inhibited the colony-forming and metastatic potential of HT1080 human fibrosarcoma cells (15). Although many studies reveal the relationship cancer mechanism and hyaluronan, there are few studies that the effect of HA synthesis inhibition and irradiation.…”

“…. cDNA was synthesized using a procedure reported previously with reaction mixture containing forward and reverse primers (Table I) (15). The reaction was carried out in a real-time PCR system (StepOne Plus; Life Technologies) under the following conditions: 95˚C for 30 sec, followed by 40 cycles of 95˚C for 5 sec, and 54˚C for 30 sec.…”