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Decreasing the ratio of matriptase/HAI‑1 by downregulation of matriptase as a potential adjuvant therapy in ovarian cancer.

Sun P, Jiang Z, Chen X, Xue L, Mao X, Ruan G, Song Y, Mustea A - Mol Med Rep (2016)

Bottom Line: The migration distance and number of invading HO‑8910PM cells were significantly increased compared with HO‑8910 cells.HO‑8910PM cells exhibited a significantly higher ratio of matriptase/HAI‑1 mRNA levels compared with HO‑8910 cells (0.51 vs. 0.24, ~2.2 fold increase).Similar increases in protein expression levels were also observed in HO‑8910PM cells compared with HO‑8910 cells.

View Article: PubMed Central - PubMed

Affiliation: Institute of Gynecologic Oncology, Department of Gynecology, Fujian Maternity and Children Health Hospital, Teaching Hospital of Fujian Medical University, Fuzhou, Fujian 350001, P.R. China.

ABSTRACT
Tumor invasion and metastasis are complex biological processes. Matriptase and its endogenous inhibitor, hepatocyte growth factor activator inhibitor‑1 (HAI‑1) are involved in invasion and metastasis. To evaluate the ratio of matriptase/HAI‑1 and their potential therapeutic value in ovarian cancer, HO‑8910 human ovarian cancer cells and the homologous high‑metastatic HO‑8910PM cells were used as in vitro cellular models ovarian cancer. The invasive and metastatic abilities, and the expression of matriptase and HAI‑1 in these cells were detected using scratch assays, Transwell chamber assays, reverse transcription‑quantitative polymerase chain reaction, western blotting and fluorescent immunocytochemistry. Following infection with lentivirus‑mediated matriptase‑targeting small interfering RNA (siRNA), cell cycle progression and apoptosis were also analyzed. The migration distance and number of invading HO‑8910PM cells were significantly increased compared with HO‑8910 cells. HO‑8910PM cells exhibited a significantly higher ratio of matriptase/HAI‑1 mRNA levels compared with HO‑8910 cells (0.51 vs. 0.24, ~2.2 fold increase). Compared with HO‑8910 cells, the matriptase mRNA level was increased by ~3.6 fold in HO‑8910PM cells, whereas the HAI‑1 mRNA level was increased by ~1.7 fold. Similar increases in protein expression levels were also observed in HO‑8910PM cells compared with HO‑8910 cells. Migration and invasiveness were positively correlated with matriptase expression level (r=0.994, P<0.01) and the ratio of matriptase/HAI‑1 (r=0.929, P<0.01). Downregulation of matriptase using siRNA resulted in inhibition of the invasive and metastatic abilities of HO‑8910PM cells, cell cycle arrest in the G0/G1 phase and increased apoptosis. The present study demonstrated that ovarian cancer cell metastasis and invasion were more dependent on upregulation of matriptase levels than downregulation of HAI‑1. Matriptase may be a potential adjuvant therapeutic target for inhibiting ovarian cancer invasion and metastasis.

No MeSH data available.


Related in: MedlinePlus

Downregulation of matriptase results in cell cycle arrest and increased apoptosis in HO-8910PM cells. (A) Flow cytometry analysis demonstrated that the percentage of cells in the G1/G0 phase was significantly higher for matriptase-depleted HO-8910PM-SI2 cells than for HO-8910PM-NC and HO-8910PM cells (42.73±0.39%, P<0.01). Tthe percentage of HO-8910PM-SI2 cells in the G2/M phase was significantly lower (4.16±0.74%) than that of HO-8910PM-NC (17.65±0.63%, P<0.01) and HO-8910PM cells (18.35±0.65%, P<0.01). Conversely, no differences in S phase content were noted among the 3 cell lines. (B) Matriptase depletion significantly decreased the percentage of surviving cells and increased the percentage of early apoptotic cells in HO-8910PM-SI2 cells compared with the negative control and wild type cells. The percentages of late apoptotic and necrotic cells were not significantly different. (C) Relative mRNA and protein expression levels of matriptase and HAI-1 were detected in HO-8910PM, HO-8910PM-SI2 and HO-8910PM-NC cells. The highest silencing efficiency was achieved in HO-8910PM-SI2 cells using matriptase-SI2. The mRNA and protein expression levels of matriptase and HAI-1 were comparable in HO-8910 and HO-8910-NC cells.#compared with HO-8910PM-NC and HO-8910PM cells, significantly fewer HO-8910PM-SI2 cells survived and the number of early apoptotic HO-8910PM-SI2 cells was significantly higher. *P>0.05 vs. HO-8910PM; #P<0.05 vs. HO-8910PM. Q1, cellular debris, Q2, late apoptotic and necrotic cells, Q3, surviving cells, Q4, early apoptotic cells. NC, negative control; SI, small interfering RNA; HAI-1, hepatocyte growth factor activator inhibitor-1.
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f5-mmr-14-02-1465: Downregulation of matriptase results in cell cycle arrest and increased apoptosis in HO-8910PM cells. (A) Flow cytometry analysis demonstrated that the percentage of cells in the G1/G0 phase was significantly higher for matriptase-depleted HO-8910PM-SI2 cells than for HO-8910PM-NC and HO-8910PM cells (42.73±0.39%, P<0.01). Tthe percentage of HO-8910PM-SI2 cells in the G2/M phase was significantly lower (4.16±0.74%) than that of HO-8910PM-NC (17.65±0.63%, P<0.01) and HO-8910PM cells (18.35±0.65%, P<0.01). Conversely, no differences in S phase content were noted among the 3 cell lines. (B) Matriptase depletion significantly decreased the percentage of surviving cells and increased the percentage of early apoptotic cells in HO-8910PM-SI2 cells compared with the negative control and wild type cells. The percentages of late apoptotic and necrotic cells were not significantly different. (C) Relative mRNA and protein expression levels of matriptase and HAI-1 were detected in HO-8910PM, HO-8910PM-SI2 and HO-8910PM-NC cells. The highest silencing efficiency was achieved in HO-8910PM-SI2 cells using matriptase-SI2. The mRNA and protein expression levels of matriptase and HAI-1 were comparable in HO-8910 and HO-8910-NC cells.#compared with HO-8910PM-NC and HO-8910PM cells, significantly fewer HO-8910PM-SI2 cells survived and the number of early apoptotic HO-8910PM-SI2 cells was significantly higher. *P>0.05 vs. HO-8910PM; #P<0.05 vs. HO-8910PM. Q1, cellular debris, Q2, late apoptotic and necrotic cells, Q3, surviving cells, Q4, early apoptotic cells. NC, negative control; SI, small interfering RNA; HAI-1, hepatocyte growth factor activator inhibitor-1.

Mentions: The percentage of G0/G1, S and G2/M phase HO-8910PM-SI2 cells were 54.81, 41.03 and 4.16%, respectively. The number of matriptase-knockdown HO-8910PM-SI2 cells in the G0/G1 phase (54.81±0.34%) was significantly increased compared with HO-8910PM-NC (43.08±0.47%) and HO-8910PM cells (42.73±0.39%; both P<0.01; Fig. 5A). Whereas, the number of G2/M phase HO-8910PM-SI2 cells (4.16±0.74%) was significantly reduced compared with HO-8910PM-NC (17.65±0.63%) and HO-8910PM (8.35±0.65%; both P<0.01). However, no significant difference in S phase content was observed between the HO-8910PM-SI2, HO-8910PM-NC and HO-8910PM cells (41.03±1.02, 39.27±0.97, and 38.92±1.12%, respectively). Furthermore, matriptase suppression decreased the percentage of surviving cells and increased the percentage of early apoptotic cells (Fig. 5B). Compared with HO-8910PM-NC (88.7±0.41%) and HO-8910PM cells (86.2±0.41%), the number surviving HO-8910PM-SI2 cells of was significantly lower (75.10±0.41%; P<0.01). Additionally, the number of early apoptotic HO-8910PM-SI2 cells (15.10±0.81%) was significantly increased compared with HO-8910PM-NC (5.2±0.39%) and HO-8910PM cells (5.3±0.33%; both P<0.01). The percentages of late apoptotic and necrotic cells were not significantly different between the cells.


Decreasing the ratio of matriptase/HAI‑1 by downregulation of matriptase as a potential adjuvant therapy in ovarian cancer.

Sun P, Jiang Z, Chen X, Xue L, Mao X, Ruan G, Song Y, Mustea A - Mol Med Rep (2016)

Downregulation of matriptase results in cell cycle arrest and increased apoptosis in HO-8910PM cells. (A) Flow cytometry analysis demonstrated that the percentage of cells in the G1/G0 phase was significantly higher for matriptase-depleted HO-8910PM-SI2 cells than for HO-8910PM-NC and HO-8910PM cells (42.73±0.39%, P<0.01). Tthe percentage of HO-8910PM-SI2 cells in the G2/M phase was significantly lower (4.16±0.74%) than that of HO-8910PM-NC (17.65±0.63%, P<0.01) and HO-8910PM cells (18.35±0.65%, P<0.01). Conversely, no differences in S phase content were noted among the 3 cell lines. (B) Matriptase depletion significantly decreased the percentage of surviving cells and increased the percentage of early apoptotic cells in HO-8910PM-SI2 cells compared with the negative control and wild type cells. The percentages of late apoptotic and necrotic cells were not significantly different. (C) Relative mRNA and protein expression levels of matriptase and HAI-1 were detected in HO-8910PM, HO-8910PM-SI2 and HO-8910PM-NC cells. The highest silencing efficiency was achieved in HO-8910PM-SI2 cells using matriptase-SI2. The mRNA and protein expression levels of matriptase and HAI-1 were comparable in HO-8910 and HO-8910-NC cells.#compared with HO-8910PM-NC and HO-8910PM cells, significantly fewer HO-8910PM-SI2 cells survived and the number of early apoptotic HO-8910PM-SI2 cells was significantly higher. *P>0.05 vs. HO-8910PM; #P<0.05 vs. HO-8910PM. Q1, cellular debris, Q2, late apoptotic and necrotic cells, Q3, surviving cells, Q4, early apoptotic cells. NC, negative control; SI, small interfering RNA; HAI-1, hepatocyte growth factor activator inhibitor-1.
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Related In: Results  -  Collection

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f5-mmr-14-02-1465: Downregulation of matriptase results in cell cycle arrest and increased apoptosis in HO-8910PM cells. (A) Flow cytometry analysis demonstrated that the percentage of cells in the G1/G0 phase was significantly higher for matriptase-depleted HO-8910PM-SI2 cells than for HO-8910PM-NC and HO-8910PM cells (42.73±0.39%, P<0.01). Tthe percentage of HO-8910PM-SI2 cells in the G2/M phase was significantly lower (4.16±0.74%) than that of HO-8910PM-NC (17.65±0.63%, P<0.01) and HO-8910PM cells (18.35±0.65%, P<0.01). Conversely, no differences in S phase content were noted among the 3 cell lines. (B) Matriptase depletion significantly decreased the percentage of surviving cells and increased the percentage of early apoptotic cells in HO-8910PM-SI2 cells compared with the negative control and wild type cells. The percentages of late apoptotic and necrotic cells were not significantly different. (C) Relative mRNA and protein expression levels of matriptase and HAI-1 were detected in HO-8910PM, HO-8910PM-SI2 and HO-8910PM-NC cells. The highest silencing efficiency was achieved in HO-8910PM-SI2 cells using matriptase-SI2. The mRNA and protein expression levels of matriptase and HAI-1 were comparable in HO-8910 and HO-8910-NC cells.#compared with HO-8910PM-NC and HO-8910PM cells, significantly fewer HO-8910PM-SI2 cells survived and the number of early apoptotic HO-8910PM-SI2 cells was significantly higher. *P>0.05 vs. HO-8910PM; #P<0.05 vs. HO-8910PM. Q1, cellular debris, Q2, late apoptotic and necrotic cells, Q3, surviving cells, Q4, early apoptotic cells. NC, negative control; SI, small interfering RNA; HAI-1, hepatocyte growth factor activator inhibitor-1.
Mentions: The percentage of G0/G1, S and G2/M phase HO-8910PM-SI2 cells were 54.81, 41.03 and 4.16%, respectively. The number of matriptase-knockdown HO-8910PM-SI2 cells in the G0/G1 phase (54.81±0.34%) was significantly increased compared with HO-8910PM-NC (43.08±0.47%) and HO-8910PM cells (42.73±0.39%; both P<0.01; Fig. 5A). Whereas, the number of G2/M phase HO-8910PM-SI2 cells (4.16±0.74%) was significantly reduced compared with HO-8910PM-NC (17.65±0.63%) and HO-8910PM (8.35±0.65%; both P<0.01). However, no significant difference in S phase content was observed between the HO-8910PM-SI2, HO-8910PM-NC and HO-8910PM cells (41.03±1.02, 39.27±0.97, and 38.92±1.12%, respectively). Furthermore, matriptase suppression decreased the percentage of surviving cells and increased the percentage of early apoptotic cells (Fig. 5B). Compared with HO-8910PM-NC (88.7±0.41%) and HO-8910PM cells (86.2±0.41%), the number surviving HO-8910PM-SI2 cells of was significantly lower (75.10±0.41%; P<0.01). Additionally, the number of early apoptotic HO-8910PM-SI2 cells (15.10±0.81%) was significantly increased compared with HO-8910PM-NC (5.2±0.39%) and HO-8910PM cells (5.3±0.33%; both P<0.01). The percentages of late apoptotic and necrotic cells were not significantly different between the cells.

Bottom Line: The migration distance and number of invading HO‑8910PM cells were significantly increased compared with HO‑8910 cells.HO‑8910PM cells exhibited a significantly higher ratio of matriptase/HAI‑1 mRNA levels compared with HO‑8910 cells (0.51 vs. 0.24, ~2.2 fold increase).Similar increases in protein expression levels were also observed in HO‑8910PM cells compared with HO‑8910 cells.

View Article: PubMed Central - PubMed

Affiliation: Institute of Gynecologic Oncology, Department of Gynecology, Fujian Maternity and Children Health Hospital, Teaching Hospital of Fujian Medical University, Fuzhou, Fujian 350001, P.R. China.

ABSTRACT
Tumor invasion and metastasis are complex biological processes. Matriptase and its endogenous inhibitor, hepatocyte growth factor activator inhibitor‑1 (HAI‑1) are involved in invasion and metastasis. To evaluate the ratio of matriptase/HAI‑1 and their potential therapeutic value in ovarian cancer, HO‑8910 human ovarian cancer cells and the homologous high‑metastatic HO‑8910PM cells were used as in vitro cellular models ovarian cancer. The invasive and metastatic abilities, and the expression of matriptase and HAI‑1 in these cells were detected using scratch assays, Transwell chamber assays, reverse transcription‑quantitative polymerase chain reaction, western blotting and fluorescent immunocytochemistry. Following infection with lentivirus‑mediated matriptase‑targeting small interfering RNA (siRNA), cell cycle progression and apoptosis were also analyzed. The migration distance and number of invading HO‑8910PM cells were significantly increased compared with HO‑8910 cells. HO‑8910PM cells exhibited a significantly higher ratio of matriptase/HAI‑1 mRNA levels compared with HO‑8910 cells (0.51 vs. 0.24, ~2.2 fold increase). Compared with HO‑8910 cells, the matriptase mRNA level was increased by ~3.6 fold in HO‑8910PM cells, whereas the HAI‑1 mRNA level was increased by ~1.7 fold. Similar increases in protein expression levels were also observed in HO‑8910PM cells compared with HO‑8910 cells. Migration and invasiveness were positively correlated with matriptase expression level (r=0.994, P<0.01) and the ratio of matriptase/HAI‑1 (r=0.929, P<0.01). Downregulation of matriptase using siRNA resulted in inhibition of the invasive and metastatic abilities of HO‑8910PM cells, cell cycle arrest in the G0/G1 phase and increased apoptosis. The present study demonstrated that ovarian cancer cell metastasis and invasion were more dependent on upregulation of matriptase levels than downregulation of HAI‑1. Matriptase may be a potential adjuvant therapeutic target for inhibiting ovarian cancer invasion and metastasis.

No MeSH data available.


Related in: MedlinePlus