Enhanced therapeutic effects for human pancreatic cancer by application K-ras and IGF-IR antisense
Yong-Mei Shen1, Xiao-Chun Yang, Chen Yang
1Radioimmunoassay Center & Clinical Laboratory, Department of MRI, the Second Affiliated Hospital of Soochow University, Suzhou 215004, Jiangsu Province, China.
Aim:
To investigate the combined effects of K-ras antisense oligodeoxynucleotide (K-ras ASODN) specific to GTT point mutation at codon 12 and type I insulin-like growth factor receptor (IGF-IR) antisense oligodeoxynucleotide (IGF-IR ASODN) on proliferation and apoptosis of human pancreatic cancer Patu8988 cells in vitro and in vivo.
Methods:
K-ras gene point mutation and its style at codon 12 of human pancreatic cancer cell line Patu8988 were detected by using polymerase chain reaction with special sequence primers (PCR-SSP) and sequence analysis. According to the mutation style, K-ras mutation ASODN specific to K-ras point mutation at codon 12 was designed and composed. After K-ras ASODN and IGF-IR ASODN treated on Patu8988 cells respectively or cooperatively, the proliferation and morphological change of Patu8988 cells were analyzed by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay, colony forming assay and transmission electron microscopy; the expression of K-ras and IGF-IR mRNA and protein in the treated cells was measured by reverse-transcript polymerase chain reaction (RT-PCR) and flow cytometry respectively; apoptosis was determined by flow cytometry. The combined antitumor activity of K-ras ASODN and IGF-IR ASODN was evaluated in BALB/c nude mice bearing human pancreatic cancer inoculated with Patu8988 cells.
Results:
The results of PCR-SSP and sequence analysis showed that the human pancreatic cancer cell line Patu8988 had point mutation at codon 12, and the mutation style was GGT-->GTT. 2-32 microg/mL K-ras ASODN and 2-32 microg/mL IGF-IR ASODN could inhibit Patu8988 cells' growth, induce apoptosis and decrease the expression of K-ras and IGF-IR mRNA and protein alone. However, there was much more effective inhibition of growth and induction of apoptosis by their combination than by each one alone. In tumor bearing mice, the combination of K-ras ASODN and IGF-IR ASODN showed a significant inhibitory effect on the growth of transplanted pancreatic cancer, resulting in a statistically significant difference compared with each alone.
Conclusion:
It has been found that K-ras ASODN combined with IGF-IR ASODN could cooperatively inhibit the growth of Patu8988 cells, and induce their apoptosis via reinforcing specific down regulation of K-ras and IGF-IR mRNA and protein expression.
Insights
Combining K-ras antisense oligodeoxynucleotide (ASODN) and type I insulin-like growth factor receptor (IGF-IR) ASODN effectively inhibits pancreatic cancer cell growth and induces apoptosis. This combination therapy enhances the downregulation of K-ras and IGF-IR expression.
Area of Science:
- Oncology
- Molecular Biology
- Genetics
Background:
- Pancreatic cancer is a leading cause of cancer-related deaths.
- The K-ras gene mutation and IGF-IR signaling are implicated in pancreatic cancer progression.
- Targeted therapies are crucial for improving treatment outcomes.
Purpose of the Study:
- To evaluate the synergistic effects of K-ras ASODN and IGF-IR ASODN on human pancreatic cancer Patu8988 cells.
- To investigate the impact of this combination therapy on cell proliferation, apoptosis, and gene expression.
- To assess the in vivo antitumor activity of K-ras ASODN and IGF-IR ASODN in a xenograft model.
Main Methods:
- K-ras mutation at codon 12 (GGT-->GTT) in Patu8988 cells was identified using PCR-SSP and sequence analysis.
- Cells were treated with K-ras ASODN and IGF-IR ASODN individually and in combination.
- Proliferation was assessed by MTT and colony formation assays; apoptosis by flow cytometry.
- K-ras and IGF-IR mRNA and protein expression were measured by RT-PCR and flow cytometry.
- Antitumor efficacy was evaluated in BALB/c nude mice.
Main Results:
- K-ras ASODN and IGF-IR ASODN alone inhibited Patu8988 cell growth and induced apoptosis.
- The combination therapy demonstrated significantly enhanced inhibition of proliferation and greater induction of apoptosis compared to monotherapy.
- Combined treatment led to a more pronounced downregulation of K-ras and IGF-IR mRNA and protein expression.
- In vivo studies showed significant tumor growth inhibition with the combination therapy.
Conclusions:
- K-ras ASODN and IGF-IR ASODN exhibit cooperative antitumor effects against pancreatic cancer cells.
- The combination therapy enhances apoptosis and inhibits proliferation by reinforcing the specific downregulation of K-ras and IGF-IR.
- This combined antisense oligodeoxynucleotide approach holds potential as a therapeutic strategy for pancreatic cancer.
Related Concept Videos
02:57Targeted Cancer Therapies
There are several types of targeted therapies against specific...
01:27Tumor Immunotherapy