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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

TPL inhibits migration and invasion of MHCC-97H cells. (A) Cell migratory ability detected by Transwell migration assay. (B) Cell migratory ability detected by wound healing assay. Data is presented as the mean ±SD; p values were calculated with the Student’s t-test.* p<0.05
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f5-medscimonit-22-1827: TPL inhibits migration and invasion of MHCC-97H cells. (A) Cell migratory ability detected by Transwell migration assay. (B) Cell migratory ability detected by wound healing assay. Data is presented as the mean ±SD; p values were calculated with the Student’s t-test.* p<0.05

Mentions: The effects of TPL on MHCC-97H cell invasion and migration were analyzed by a wound-healing assay and by Transwell Boyden chamber assay. In the wound-healing assay, the results showed that the area change for wound-healing in the TPL treated MHCC-97H cells was reduced compared to the vehicle-treated cells (p<0.05) (Figure 5A). In the Transwell Matrigel invasion assays, the results showed that the number of cells in the lower chamber of the Transwell was significantly decreased in TPL treated MHCC-97H cells compared to the vehicle-treated cells (p<0.05) (Figures 5B). Taken together, these results indicated that TPL inhibits both the migration and invasion of 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)

TPL inhibits migration and invasion of MHCC-97H cells. (A) Cell migratory ability detected by Transwell migration assay. (B) Cell migratory ability detected by wound healing assay. Data is presented as the mean ±SD; p values were calculated with the Student’s t-test.* p<0.05
© Copyright Policy - open-access
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC4920093&req=5

f5-medscimonit-22-1827: TPL inhibits migration and invasion of MHCC-97H cells. (A) Cell migratory ability detected by Transwell migration assay. (B) Cell migratory ability detected by wound healing assay. Data is presented as the mean ±SD; p values were calculated with the Student’s t-test.* p<0.05
Mentions: The effects of TPL on MHCC-97H cell invasion and migration were analyzed by a wound-healing assay and by Transwell Boyden chamber assay. In the wound-healing assay, the results showed that the area change for wound-healing in the TPL treated MHCC-97H cells was reduced compared to the vehicle-treated cells (p<0.05) (Figure 5A). In the Transwell Matrigel invasion assays, the results showed that the number of cells in the lower chamber of the Transwell was significantly decreased in TPL treated MHCC-97H cells compared to the vehicle-treated cells (p<0.05) (Figures 5B). Taken together, these results indicated that TPL inhibits both the migration and invasion of 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