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Efficient Inhibition of Ovarian Cancer by Gelonin Toxin Gene Delivered by Biodegradable Cationic Heparin-polyethyleneimine Nanogels.

Bai Y, Gou M, Yi T, Yang L, Liu L, Lin X, Su D, Wei Y, Zhao X - Int J Med Sci (2015)

Bottom Line: Treatment for intraperitoneal carcinomatosis with pGelonin/HPEI complexes reduced the tumor weight by ~58.55% compared to the control groups (P<0.05).The antitumor effect was accompanied by increased apoptosis and reduced cell proliferation (P<0.05).No significant side effects were observed with i.p. administration of the pGelonin/HPEI complexes.

View Article: PubMed Central - PubMed

Affiliation: 1. Department of Gynecology and Obstetrics, Key Laboratory of Obstetric and Gynecologic and Pediatric Diseases and Birth Defects of Ministry of Education, West China Second Hospital, Sichuan University, Chengdu, Sichuan 610041, P.R. China.

ABSTRACT
The use of toxins for cancer therapy has great promise. Gelonin, a potent plant toxin, causes cell death by inactivating the 60S ribosomal subunit. Recently, we developed a novel gene delivery system using biodegradable cationic heparin-polyethyleneimine (HPEI) nanogels. In the current study, the antitumor activity of a recombinant plasmid expressing gelonin (pGelonin) on human ovarian cancer was assessed. The application of HPEI nanogels, was also evaluated. Gelonin-cDNA was cloned into the pVAX1 plasmid vector and transfected into SKOV3 human ovarian cancer cells using biodegradable cationic HPEI nanogels. The expression of gelonin in vitro and in vivo was confirmed using RT-PCR and western blot analysis. Cell viability and apoptosis were examined using an MTT assay and flow cytometric analysis. For the in vivo study, an SKOV3 intraperitoneal ovarian carcinomatosis model was established, and nude mice were randomly assigned into four groups receiving i.p. administration of pGelonin/HPEI complexes, pVAX/HPEI complexes, HPEI alone and 5% glucose solution. The tumor weight was monitored, and a TUNEL assay and Ki-67 immunohistochemistry were performed to evaluate apoptosis and cell proliferation in the tumor tissue sections, respectively. Gelonin was efficiently expressed in SKOV3 cancer cells in vitro and in vivo using pGelonin incorporated with HPEI nanogels. The pGelonin/HPEI complexes inhibited cell viability and induced apoptosis in the cell culture. Treatment for intraperitoneal carcinomatosis with pGelonin/HPEI complexes reduced the tumor weight by ~58.55% compared to the control groups (P<0.05). The antitumor effect was accompanied by increased apoptosis and reduced cell proliferation (P<0.05). No significant side effects were observed with i.p. administration of the pGelonin/HPEI complexes. Our data indicate that HPEI nanogel-delivered pGelonin may have promising applications against human ovarian cancer.

No MeSH data available.


Related in: MedlinePlus

Expression of gelonin in intraperitoneal tumor tissues. (A) Expression of gelonin in intraperitoneal tumor tissues detected by RT-PCR (left) and western blot analysis (right). Gelonin mRNA and protein were detected in pGelonin/HPEI group, by contrast, no expression of gelonin was observed in medium alone, HPEI or pVAX/HPEI group. GAPDH served as internal control. (B) immunostaining of gelonin in intraperitoneal tumor tissues. Original magnification, ×400.
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Figure 3: Expression of gelonin in intraperitoneal tumor tissues. (A) Expression of gelonin in intraperitoneal tumor tissues detected by RT-PCR (left) and western blot analysis (right). Gelonin mRNA and protein were detected in pGelonin/HPEI group, by contrast, no expression of gelonin was observed in medium alone, HPEI or pVAX/HPEI group. GAPDH served as internal control. (B) immunostaining of gelonin in intraperitoneal tumor tissues. Original magnification, ×400.

Mentions: To examine the expression of gelonin in vivo, an intraperitoneal ovarian carcinomatosis model was established and nude mice were treated with pGelonin/HPEI complexes, pVAX/HPEI complexes, HPEI nanogels or medium alone. At 3 days after the final treatment, intraperitoneal tumors were collected for RT-PCR, western blot analysis and immunostaining. The expression of gelonin mRNA and protein were detected in tumor tissues from the pGelonin/HPEI complexes-treated group, whereas the pVAX/HPEI complexes, HPEI nanogels or medium groups had no expression of gelonin (Fig. 3A). Furthermore, immunostaining of gelonin in tumor tissues also suggested expression of gelonin in the pGelonin/HPEI complexes-treated group as shown in Fig. 3B.


Efficient Inhibition of Ovarian Cancer by Gelonin Toxin Gene Delivered by Biodegradable Cationic Heparin-polyethyleneimine Nanogels.

Bai Y, Gou M, Yi T, Yang L, Liu L, Lin X, Su D, Wei Y, Zhao X - Int J Med Sci (2015)

Expression of gelonin in intraperitoneal tumor tissues. (A) Expression of gelonin in intraperitoneal tumor tissues detected by RT-PCR (left) and western blot analysis (right). Gelonin mRNA and protein were detected in pGelonin/HPEI group, by contrast, no expression of gelonin was observed in medium alone, HPEI or pVAX/HPEI group. GAPDH served as internal control. (B) immunostaining of gelonin in intraperitoneal tumor tissues. Original magnification, ×400.
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Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC4441064&req=5

Figure 3: Expression of gelonin in intraperitoneal tumor tissues. (A) Expression of gelonin in intraperitoneal tumor tissues detected by RT-PCR (left) and western blot analysis (right). Gelonin mRNA and protein were detected in pGelonin/HPEI group, by contrast, no expression of gelonin was observed in medium alone, HPEI or pVAX/HPEI group. GAPDH served as internal control. (B) immunostaining of gelonin in intraperitoneal tumor tissues. Original magnification, ×400.
Mentions: To examine the expression of gelonin in vivo, an intraperitoneal ovarian carcinomatosis model was established and nude mice were treated with pGelonin/HPEI complexes, pVAX/HPEI complexes, HPEI nanogels or medium alone. At 3 days after the final treatment, intraperitoneal tumors were collected for RT-PCR, western blot analysis and immunostaining. The expression of gelonin mRNA and protein were detected in tumor tissues from the pGelonin/HPEI complexes-treated group, whereas the pVAX/HPEI complexes, HPEI nanogels or medium groups had no expression of gelonin (Fig. 3A). Furthermore, immunostaining of gelonin in tumor tissues also suggested expression of gelonin in the pGelonin/HPEI complexes-treated group as shown in Fig. 3B.

Bottom Line: Treatment for intraperitoneal carcinomatosis with pGelonin/HPEI complexes reduced the tumor weight by ~58.55% compared to the control groups (P<0.05).The antitumor effect was accompanied by increased apoptosis and reduced cell proliferation (P<0.05).No significant side effects were observed with i.p. administration of the pGelonin/HPEI complexes.

View Article: PubMed Central - PubMed

Affiliation: 1. Department of Gynecology and Obstetrics, Key Laboratory of Obstetric and Gynecologic and Pediatric Diseases and Birth Defects of Ministry of Education, West China Second Hospital, Sichuan University, Chengdu, Sichuan 610041, P.R. China.

ABSTRACT
The use of toxins for cancer therapy has great promise. Gelonin, a potent plant toxin, causes cell death by inactivating the 60S ribosomal subunit. Recently, we developed a novel gene delivery system using biodegradable cationic heparin-polyethyleneimine (HPEI) nanogels. In the current study, the antitumor activity of a recombinant plasmid expressing gelonin (pGelonin) on human ovarian cancer was assessed. The application of HPEI nanogels, was also evaluated. Gelonin-cDNA was cloned into the pVAX1 plasmid vector and transfected into SKOV3 human ovarian cancer cells using biodegradable cationic HPEI nanogels. The expression of gelonin in vitro and in vivo was confirmed using RT-PCR and western blot analysis. Cell viability and apoptosis were examined using an MTT assay and flow cytometric analysis. For the in vivo study, an SKOV3 intraperitoneal ovarian carcinomatosis model was established, and nude mice were randomly assigned into four groups receiving i.p. administration of pGelonin/HPEI complexes, pVAX/HPEI complexes, HPEI alone and 5% glucose solution. The tumor weight was monitored, and a TUNEL assay and Ki-67 immunohistochemistry were performed to evaluate apoptosis and cell proliferation in the tumor tissue sections, respectively. Gelonin was efficiently expressed in SKOV3 cancer cells in vitro and in vivo using pGelonin incorporated with HPEI nanogels. The pGelonin/HPEI complexes inhibited cell viability and induced apoptosis in the cell culture. Treatment for intraperitoneal carcinomatosis with pGelonin/HPEI complexes reduced the tumor weight by ~58.55% compared to the control groups (P<0.05). The antitumor effect was accompanied by increased apoptosis and reduced cell proliferation (P<0.05). No significant side effects were observed with i.p. administration of the pGelonin/HPEI complexes. Our data indicate that HPEI nanogel-delivered pGelonin may have promising applications against human ovarian cancer.

No MeSH data available.


Related in: MedlinePlus