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Triptolide Inhibits Invasion and Tumorigenesis of Hepatocellular Carcinoma MHCC-97H Cells Through NF-κB Signaling.

Wang H, Ma D, Wang C, Zhao S, Liu C - Med. Sci. Monit. (2016)

Bottom Line: To detect the effect of NF-κB on TPL-induced signal pathways, MHCC-97H cells were transfected with p65 siRNA or p65 cDNA, then treated with TPL.RESULTS We showed that TPL treatment significantly suppressed growth and induced apoptosis of MHCC-97H cells in a dose- and time-dependent manner in vitro.Overexpression of p65 restored MMP-9 activity and inhibited the TPL anti-tumor effect on MHCC-97H cells.

View Article: PubMed Central - PubMed

Affiliation: Department of Radiation Oncology, The Affiliated Hospital of Qingdao University, Qingdao, Shangdong, China (mainland).

ABSTRACT
BACKGROUND We investigated whether the plant-derived agent triptolide (TPL) could effectively inhibit the growth and invasion of human hepatocellular carcinoma (HCC) cells. MATERIAL AND METHODS MHCC-97H cells were treated with various concentration of TPL for various times. To detect the effect of NF-κB on TPL-induced signal pathways, MHCC-97H cells were transfected with p65 siRNA or p65 cDNA, then treated with TPL. We detected cell survival and apoptosis by MTT, soft-agar colony formation assay, flow cytometry, and TUNEL assay. Cell migration and invasion was determined by Matrigel invasion and a wound-healing assay. NF-κB activity was detected by electrophoretic mobility shift assay (EMSA); MMP-9 activity was detected by ELISA. Western blot and real-time PCR (RT-PCR) assays were used to detect p65 and MMP-9 protein and mRNA expression. A subcutaneously implanted tumor model of MHCC-97H cells in nude mice was used to assess the effects of TPL on tumorigenesis in vivo. RESULTS We showed that TPL treatment significantly suppressed growth and induced apoptosis of MHCC-97H cells in a dose- and time-dependent manner in vitro. Furthermore, TPL treatment inhibited invasion in vitro and inhibited the growth and lung metastasis of MHCC-97H cells in vivo. NF-κB and MMP-9 were inactivated with TPL treatment. Overexpression of p65 restored MMP-9 activity and inhibited the TPL anti-tumor effect on MHCC-97H cells. Knockdown of p65 blocked MMP-9 activation and enhanced TPL-induced cell apoptosis and survival inhibition, and TPL inhibition of migration and invasion in vitro. CONCLUSIONS TPL treatment inhibited MHCC-97H cell growth, invasion, and metastasis in vitro and vivo, suggesting that TPL could be developed as a potential therapeutic agent for the treatment of HCC.

No MeSH data available.


Related in: MedlinePlus

Effect of TPL on MHCC-97H cell growth and apoptosis. (A) Dose and time responses of growth of MHCC-97H cells with TPL treatment. Cells were seeded in 96-well plates at 5,000 cells per well and treated with varied concentrations of TPL or for different time periods. After treatment, cell densities were determined by MTT assay. Each value represents the mean ±SD (n=5) of 3 independent experiments. * p<0.05, compared to the control. (B) Cell death assay for measuring apoptosis induced by TPL. Apoptosis was measured by flow cytometry. Values are reported as mean ±SD. * p<0.05, compared to the control. Cell apoptosis was also detected by TUNEL assay. Values are reported as mean ±SD. * p<0.05, compared to the control.
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f1-medscimonit-22-1827: Effect of TPL on MHCC-97H cell growth and apoptosis. (A) Dose and time responses of growth of MHCC-97H cells with TPL treatment. Cells were seeded in 96-well plates at 5,000 cells per well and treated with varied concentrations of TPL or for different time periods. After treatment, cell densities were determined by MTT assay. Each value represents the mean ±SD (n=5) of 3 independent experiments. * p<0.05, compared to the control. (B) Cell death assay for measuring apoptosis induced by TPL. Apoptosis was measured by flow cytometry. Values are reported as mean ±SD. * p<0.05, compared to the control. Cell apoptosis was also detected by TUNEL assay. Values are reported as mean ±SD. * p<0.05, compared to the control.

Mentions: MHCC-97H cells was treated with 5, 15, or 25 μM TPL for 24–72 hours. As shown in Figure 1A, TPL treatment resulted in cell growth inhibition in a dose- and time-dependent manner in MHCC-97H cells by MTT assay. The induction of apoptosis by TPL in MHCC-97H cells was also found in a dose-dependent manner by flow cytometry assay (Figure 1B) and TUNEL assay (Figure 1C). These results provided convincing data that TPL induced apoptosis, resulting in cell growth inhibition in MHCC-97H cells.


Triptolide Inhibits Invasion and Tumorigenesis of Hepatocellular Carcinoma MHCC-97H Cells Through NF-κB Signaling.

Wang H, Ma D, Wang C, Zhao S, Liu C - Med. Sci. Monit. (2016)

Effect of TPL on MHCC-97H cell growth and apoptosis. (A) Dose and time responses of growth of MHCC-97H cells with TPL treatment. Cells were seeded in 96-well plates at 5,000 cells per well and treated with varied concentrations of TPL or for different time periods. After treatment, cell densities were determined by MTT assay. Each value represents the mean ±SD (n=5) of 3 independent experiments. * p<0.05, compared to the control. (B) Cell death assay for measuring apoptosis induced by TPL. Apoptosis was measured by flow cytometry. Values are reported as mean ±SD. * p<0.05, compared to the control. Cell apoptosis was also detected by TUNEL assay. Values are reported as mean ±SD. * p<0.05, compared to the control.
© Copyright Policy - open-access
Related In: Results  -  Collection

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getmorefigures.php?uid=PMC4920093&req=5

f1-medscimonit-22-1827: Effect of TPL on MHCC-97H cell growth and apoptosis. (A) Dose and time responses of growth of MHCC-97H cells with TPL treatment. Cells were seeded in 96-well plates at 5,000 cells per well and treated with varied concentrations of TPL or for different time periods. After treatment, cell densities were determined by MTT assay. Each value represents the mean ±SD (n=5) of 3 independent experiments. * p<0.05, compared to the control. (B) Cell death assay for measuring apoptosis induced by TPL. Apoptosis was measured by flow cytometry. Values are reported as mean ±SD. * p<0.05, compared to the control. Cell apoptosis was also detected by TUNEL assay. Values are reported as mean ±SD. * p<0.05, compared to the control.
Mentions: MHCC-97H cells was treated with 5, 15, or 25 μM TPL for 24–72 hours. As shown in Figure 1A, TPL treatment resulted in cell growth inhibition in a dose- and time-dependent manner in MHCC-97H cells by MTT assay. The induction of apoptosis by TPL in MHCC-97H cells was also found in a dose-dependent manner by flow cytometry assay (Figure 1B) and TUNEL assay (Figure 1C). These results provided convincing data that TPL induced apoptosis, resulting in cell growth inhibition in MHCC-97H cells.

Bottom Line: To detect the effect of NF-κB on TPL-induced signal pathways, MHCC-97H cells were transfected with p65 siRNA or p65 cDNA, then treated with TPL.RESULTS We showed that TPL treatment significantly suppressed growth and induced apoptosis of MHCC-97H cells in a dose- and time-dependent manner in vitro.Overexpression of p65 restored MMP-9 activity and inhibited the TPL anti-tumor effect on MHCC-97H cells.

View Article: PubMed Central - PubMed

Affiliation: Department of Radiation Oncology, The Affiliated Hospital of Qingdao University, Qingdao, Shangdong, China (mainland).

ABSTRACT
BACKGROUND We investigated whether the plant-derived agent triptolide (TPL) could effectively inhibit the growth and invasion of human hepatocellular carcinoma (HCC) cells. MATERIAL AND METHODS MHCC-97H cells were treated with various concentration of TPL for various times. To detect the effect of NF-κB on TPL-induced signal pathways, MHCC-97H cells were transfected with p65 siRNA or p65 cDNA, then treated with TPL. We detected cell survival and apoptosis by MTT, soft-agar colony formation assay, flow cytometry, and TUNEL assay. Cell migration and invasion was determined by Matrigel invasion and a wound-healing assay. NF-κB activity was detected by electrophoretic mobility shift assay (EMSA); MMP-9 activity was detected by ELISA. Western blot and real-time PCR (RT-PCR) assays were used to detect p65 and MMP-9 protein and mRNA expression. A subcutaneously implanted tumor model of MHCC-97H cells in nude mice was used to assess the effects of TPL on tumorigenesis in vivo. RESULTS We showed that TPL treatment significantly suppressed growth and induced apoptosis of MHCC-97H cells in a dose- and time-dependent manner in vitro. Furthermore, TPL treatment inhibited invasion in vitro and inhibited the growth and lung metastasis of MHCC-97H cells in vivo. NF-κB and MMP-9 were inactivated with TPL treatment. Overexpression of p65 restored MMP-9 activity and inhibited the TPL anti-tumor effect on MHCC-97H cells. Knockdown of p65 blocked MMP-9 activation and enhanced TPL-induced cell apoptosis and survival inhibition, and TPL inhibition of migration and invasion in vitro. CONCLUSIONS TPL treatment inhibited MHCC-97H cell growth, invasion, and metastasis in vitro and vivo, suggesting that TPL could be developed as a potential therapeutic agent for the treatment of HCC.

No MeSH data available.


Related in: MedlinePlus