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Gene silencing of heparanase results in suppression of invasion and migration of hepatoma cells.

Dong W, Zhao H, Zhang C, Geng P - World J Surg Oncol (2014)

Bottom Line: At 72 h post-transfection, the expression of the PTGS group of genes had recovered unlike the TGS group.At 96 h post-transfection, the expression of the heparanase gene had recovered in both the TGS group and PTGS group.TGS can effectively interfere with the heparanase gene to reduce the invasion and migration of hepatoma SMCC-7721 cells.

View Article: PubMed Central - HTML - PubMed

Affiliation: Department of Oncology, First Affiliated Hospital, Chinese PLA General Hospital, Beijing 100048, China. w_hxiao@hotmail.com.

ABSTRACT

Background: This study investigated the effect of transcriptional gene silencing (TGS) of the heparanase gene on hepatoma SMCC-7721 cells.

Methods: SiRNAs targeting the promoter region and coding region of the heparanase gene were designed and synthesized. Then the siRNAs were transfected into hepatoma SMCC-7721 cells by nuclear transfection or cytoplasmic transfection. The expression of heparanase was detected by RT-PCR and Western blotting 48 h, 72 h and 96 h post-transfection. In addition, wound healing and invasion assays were performed to estimate the effect of TGS of the heparanase gene on the migration and invasion of hepatoma SMCC-7721 cells.

Results: Protein and mRNA expression of the heparanase gene were interfered with by TGS or post-transcriptional gene silencing (PTGS) 48 h after transfection. At 72 h post-transfection, the expression of the PTGS group of genes had recovered unlike the TGS group. At 96 h post-transfection, the expression of the heparanase gene had recovered in both the TGS group and PTGS group. Invasion and wound healing assays showed that both TGS and PTGS of the heparanase gene could inhibit invasion and migration of hepatoma SMCC-7721 cells, especially the TGS group.

Conclusions: TGS can effectively interfere with the heparanase gene to reduce the invasion and migration of hepatoma SMCC-7721 cells.

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Related in: MedlinePlus

Protein expression of heparanase in TGS and PTGS groups. (A) Protein expression of heparanase in the TGS and PTGS groups. (B) Semi-quantitation of heparanase protein in the TGS and PTGS groups. control: untransfected cells; PTGS, post-transcriptional gene silencing; PTGS48h: 48 h after PTGS transfection; PTGS72h: 72 h after PTGS transfection; PTGS96h: 96 h after TGS transfection; TGS, transcriptional gene silencing; TGS48h: 48 h after TGS transfection; TGS72h: 72 h after TGS transfection; TGS96h: 96 h after TGS transfection. HPA, heparanase.
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Figure 2: Protein expression of heparanase in TGS and PTGS groups. (A) Protein expression of heparanase in the TGS and PTGS groups. (B) Semi-quantitation of heparanase protein in the TGS and PTGS groups. control: untransfected cells; PTGS, post-transcriptional gene silencing; PTGS48h: 48 h after PTGS transfection; PTGS72h: 72 h after PTGS transfection; PTGS96h: 96 h after TGS transfection; TGS, transcriptional gene silencing; TGS48h: 48 h after TGS transfection; TGS72h: 72 h after TGS transfection; TGS96h: 96 h after TGS transfection. HPA, heparanase.

Mentions: The expression of the heparanase protein was assessed using Western blotting and the results are shown in Figure 2. The heparanase expression of the TGS group was successfully interfered with at 48 h and 72 h post-transfection. The heparanase expression of the PTGS group had recovered at 72 h post-transfection. As with gene expression, the heparanase protein levels of both TGS and PTGS transfected hepatoma SMCC-7721 cells had recovered at 96 h post-transfection. These findings are consistent with those for mRNA expression.


Gene silencing of heparanase results in suppression of invasion and migration of hepatoma cells.

Dong W, Zhao H, Zhang C, Geng P - World J Surg Oncol (2014)

Protein expression of heparanase in TGS and PTGS groups. (A) Protein expression of heparanase in the TGS and PTGS groups. (B) Semi-quantitation of heparanase protein in the TGS and PTGS groups. control: untransfected cells; PTGS, post-transcriptional gene silencing; PTGS48h: 48 h after PTGS transfection; PTGS72h: 72 h after PTGS transfection; PTGS96h: 96 h after TGS transfection; TGS, transcriptional gene silencing; TGS48h: 48 h after TGS transfection; TGS72h: 72 h after TGS transfection; TGS96h: 96 h after TGS transfection. HPA, heparanase.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4222455&req=5

Figure 2: Protein expression of heparanase in TGS and PTGS groups. (A) Protein expression of heparanase in the TGS and PTGS groups. (B) Semi-quantitation of heparanase protein in the TGS and PTGS groups. control: untransfected cells; PTGS, post-transcriptional gene silencing; PTGS48h: 48 h after PTGS transfection; PTGS72h: 72 h after PTGS transfection; PTGS96h: 96 h after TGS transfection; TGS, transcriptional gene silencing; TGS48h: 48 h after TGS transfection; TGS72h: 72 h after TGS transfection; TGS96h: 96 h after TGS transfection. HPA, heparanase.
Mentions: The expression of the heparanase protein was assessed using Western blotting and the results are shown in Figure 2. The heparanase expression of the TGS group was successfully interfered with at 48 h and 72 h post-transfection. The heparanase expression of the PTGS group had recovered at 72 h post-transfection. As with gene expression, the heparanase protein levels of both TGS and PTGS transfected hepatoma SMCC-7721 cells had recovered at 96 h post-transfection. These findings are consistent with those for mRNA expression.

Bottom Line: At 72 h post-transfection, the expression of the PTGS group of genes had recovered unlike the TGS group.At 96 h post-transfection, the expression of the heparanase gene had recovered in both the TGS group and PTGS group.TGS can effectively interfere with the heparanase gene to reduce the invasion and migration of hepatoma SMCC-7721 cells.

View Article: PubMed Central - HTML - PubMed

Affiliation: Department of Oncology, First Affiliated Hospital, Chinese PLA General Hospital, Beijing 100048, China. w_hxiao@hotmail.com.

ABSTRACT

Background: This study investigated the effect of transcriptional gene silencing (TGS) of the heparanase gene on hepatoma SMCC-7721 cells.

Methods: SiRNAs targeting the promoter region and coding region of the heparanase gene were designed and synthesized. Then the siRNAs were transfected into hepatoma SMCC-7721 cells by nuclear transfection or cytoplasmic transfection. The expression of heparanase was detected by RT-PCR and Western blotting 48 h, 72 h and 96 h post-transfection. In addition, wound healing and invasion assays were performed to estimate the effect of TGS of the heparanase gene on the migration and invasion of hepatoma SMCC-7721 cells.

Results: Protein and mRNA expression of the heparanase gene were interfered with by TGS or post-transcriptional gene silencing (PTGS) 48 h after transfection. At 72 h post-transfection, the expression of the PTGS group of genes had recovered unlike the TGS group. At 96 h post-transfection, the expression of the heparanase gene had recovered in both the TGS group and PTGS group. Invasion and wound healing assays showed that both TGS and PTGS of the heparanase gene could inhibit invasion and migration of hepatoma SMCC-7721 cells, especially the TGS group.

Conclusions: TGS can effectively interfere with the heparanase gene to reduce the invasion and migration of hepatoma SMCC-7721 cells.

Show MeSH
Related in: MedlinePlus