Objective: To study the therapeutic effect and mechanisms of recombinant adenovirus Ad-p14ARF in hepatocel- lular carcinoma cell lines. Methods: Morphology and trypan blue assay were adopted to evaluate the proliferat...Objective: To study the therapeutic effect and mechanisms of recombinant adenovirus Ad-p14ARF in hepatocel- lular carcinoma cell lines. Methods: Morphology and trypan blue assay were adopted to evaluate the proliferation of different liver cancer cells after Ad-p14ARF infection. Cell apoptosis was confirmed by detecting phosphatidylserine (PS) externaliza- tion with Annexin V/PI double staining. Western blotting assay analyzed the expression of related proteins. Subcutaneous tumor model of BEL7402 was established to evaluate the therapeutic ability of Ad-p14ARF. Results: Ad-p14ARF suppressed cell growth, proliferation and promoted cell apoptosis of cancer cell lines with different genetic background. Ad-p14ARF in- hibited growth of liver cancer cells (HepG2, BEL7402) in a dose-dependent manner. Ad-p14ARF leaded to overexpression of Bax and p21, which were the downstream regulating genes of p53. Ad-p14ARF suppressed tumor growth significantly in the experimental therapy in nude mice bearing subcutaneous tumor of BEL7402. Conclusion: P14ARF gene is a powerful tumor suppressor gene to be used in cancer gene therapy. It may play an important role in gene therapy against the malignancies in the future.展开更多
Objective To investigate the relationship between alterations of p16 INK4a and p14 ARF genes and gastric carcinogenesis Methods The tumors and neighboring gastric tissues from 48 patients with gastric ca...Objective To investigate the relationship between alterations of p16 INK4a and p14 ARF genes and gastric carcinogenesis Methods The tumors and neighboring gastric tissues from 48 patients with gastric cancer were studied The homozygous deletion, mutation, methylation of the CpG islands, and mRNA expression of p16 INK4a and p14 ARF genes were assessed by PCR, PCR SSCP, PCR based methylation assay, and RT PCR Results ① The homozygous deletion rate of p16 INK4a and p14 ARF was 35 4% (17/48), and no homozygous deletion was examined in any gastric tissue neighboring the tumor ② There was no point mutation of p16 INK4a and p14 ARF in 31 gastric cancers without homozygous deletion or in the matched gastric tissues adjacent to the tumor ③ Methylation of the CpG islands of p16 INK4a and p14 ARF was detected in 47 9% (23/48) of gastric cancers, while methylation was observed only in 2 of 48 gastric tissues neighboring the cancer with a significant difference ( P <0 01) ④ The loss rate of p16 INK4a mRNA was 47 9% (23/48) in gastric cancer, and the patients of the combined methylation of exons 1α and 2 had a higher loss rate (100%, 6/6) of p16 INK4a mRNA than those of the methylation of the other exons (11 8%, 2/17, P <0 01); the loss rate of p14 ARF mRNA was 45 8%(22/48) in gastric cancer, and patients with the combined methylation of exons 1β and 2 had a higher loss rate (100%, 3/3) of p14 ARF mRNA than those of the methylation of the other exons (15%, 3/20, P <0 05) ⑤ The combined loss of p16 INK4a and p14 ARF mRNAs was examined in 1 (5 6%) of 18 patients of well and moderately differentiated carcinomas, and 11 (36 7%) of 30 patients of poorly and not differentiated carcinomas with a significant difference ( P <0 05) Conclusion p16 INK4a and p14 ARF genes are frequently inactivated by homozygous deletion and methylation of the 5'CpG islands in gastric cancer, which may play an important role in the carcinogenesis of gastric cancer展开更多
基金Supported by the National Key Basic Research Program (NKBRP, 973 program, No. 2002CB513100-8).
文摘Objective: To study the therapeutic effect and mechanisms of recombinant adenovirus Ad-p14ARF in hepatocel- lular carcinoma cell lines. Methods: Morphology and trypan blue assay were adopted to evaluate the proliferation of different liver cancer cells after Ad-p14ARF infection. Cell apoptosis was confirmed by detecting phosphatidylserine (PS) externaliza- tion with Annexin V/PI double staining. Western blotting assay analyzed the expression of related proteins. Subcutaneous tumor model of BEL7402 was established to evaluate the therapeutic ability of Ad-p14ARF. Results: Ad-p14ARF suppressed cell growth, proliferation and promoted cell apoptosis of cancer cell lines with different genetic background. Ad-p14ARF in- hibited growth of liver cancer cells (HepG2, BEL7402) in a dose-dependent manner. Ad-p14ARF leaded to overexpression of Bax and p21, which were the downstream regulating genes of p53. Ad-p14ARF suppressed tumor growth significantly in the experimental therapy in nude mice bearing subcutaneous tumor of BEL7402. Conclusion: P14ARF gene is a powerful tumor suppressor gene to be used in cancer gene therapy. It may play an important role in gene therapy against the malignancies in the future.
文摘Objective To investigate the relationship between alterations of p16 INK4a and p14 ARF genes and gastric carcinogenesis Methods The tumors and neighboring gastric tissues from 48 patients with gastric cancer were studied The homozygous deletion, mutation, methylation of the CpG islands, and mRNA expression of p16 INK4a and p14 ARF genes were assessed by PCR, PCR SSCP, PCR based methylation assay, and RT PCR Results ① The homozygous deletion rate of p16 INK4a and p14 ARF was 35 4% (17/48), and no homozygous deletion was examined in any gastric tissue neighboring the tumor ② There was no point mutation of p16 INK4a and p14 ARF in 31 gastric cancers without homozygous deletion or in the matched gastric tissues adjacent to the tumor ③ Methylation of the CpG islands of p16 INK4a and p14 ARF was detected in 47 9% (23/48) of gastric cancers, while methylation was observed only in 2 of 48 gastric tissues neighboring the cancer with a significant difference ( P <0 01) ④ The loss rate of p16 INK4a mRNA was 47 9% (23/48) in gastric cancer, and the patients of the combined methylation of exons 1α and 2 had a higher loss rate (100%, 6/6) of p16 INK4a mRNA than those of the methylation of the other exons (11 8%, 2/17, P <0 01); the loss rate of p14 ARF mRNA was 45 8%(22/48) in gastric cancer, and patients with the combined methylation of exons 1β and 2 had a higher loss rate (100%, 3/3) of p14 ARF mRNA than those of the methylation of the other exons (15%, 3/20, P <0 05) ⑤ The combined loss of p16 INK4a and p14 ARF mRNAs was examined in 1 (5 6%) of 18 patients of well and moderately differentiated carcinomas, and 11 (36 7%) of 30 patients of poorly and not differentiated carcinomas with a significant difference ( P <0 05) Conclusion p16 INK4a and p14 ARF genes are frequently inactivated by homozygous deletion and methylation of the 5'CpG islands in gastric cancer, which may play an important role in the carcinogenesis of gastric cancer
