[Effect of arsenic trioxide on the expression of cyclins gene in HL60 cells].

OBJECTIVE: To investigate the expression of cyclins related gene in HL60 cells before and after arsenic trioxide treatment by gene chip. METHODS: Total mRNAs were extracted from untreated or arsenic trioxide treated HL60 cells. Then two cDNA probes were made from these two mRNAs which were labeled by fluoro link Cy3-ducpp deoxyuridine triphosphate (Cy3-dUTP) or Cy5-dUTP fluorescence dyes respectively, hybridized with gene chip and scanned for fluorescent intensity. Different expression genes were then screened out. Cell apoptosis was detected by electron microscopy, in site cell apoptosis detection kit, DNA agarose gel electrophoresis and flow cytometry (FCM). RESULTS: Among the 82 genes which expressed differently after treatment with arsenic trioxide, 34 genes were up-regulated, while 48 genes down-regulated. It was detected that 15 micro mol/L As(2)O(3) can definitively induce HL60 cells to go apoptosis by FCM. Rate of apoptotic HL60 cells in control group is 1.7%, and As(2)O(3) group is 26.1%. CONCLUSION: Cyclin B1, proliferating cell nuclear antigen (PCNA), insulin like growth factor binding protein (IGFBP) et al may play an important role in HL60 cell apoptosis induced by arsenic trioxide.

Association of NFKBIA polymorphism with colorectal cancer risk and prognosis in Swedish and Chinese populations.

OBJECTIVE: The inhibitory proteins, IkappaBs, regulate the activity of nuclear factor kappa-beta (NF-kappaB), which is implicated in tumorigenesis by regulating expression of a variety of genes involved in cellular transformation, proliferation, invasion, angiogenesis and metastasis. Variants in the genes encoding IkappaBs may be involved in cancer development through the activation of NF-kappaB. The objective of this study was to investigate the susceptibility of an A to G variation (rs696) in the 3' UTR of NFKBIA (encoding IkappaBalpha) to colorectal cancer (CRC) and the association of this polymorphism with clinicopathologic variables in CRC patients. MATERIAL AND METHODS: A case-control study was carried out on a Swedish (155 CRCs, 438 controls) and a Chinese population (199 CRCs, 577 controls). The genotype of NFKBIA was determined by PCR-restriction fragment length polymorphism. RESULTS: The frequency of the AG genotype was increased in the Chinese patients >or=50 years of age compared with the Chinese controls (odds ratio (OR)=3.06, 95% confidence interval (CI)=1.55-6.02, p=0.001), even when adjusted for age (OR=3.20, 95% CI=1.61-6.38, p=0.001). The GG genotype of NFKBIA was related to a poorer survival rate in the Swedish patients, independent of gender, age, tumour location, Dukes' stage and differentiation (hazard ratio = 3.10, 95% Cl=1.28-7.60, p=0.01). CONCLUSIONS: Chinese individuals >or=50 years of age carrying the AG genotype of NFKBIA may be at an increased risk of developing CRC, and the GG genotype of NFKBIA may be considered as a prognostic factor for Swedish CRC patients.

[Effect of cyclin G1 antisense oligodeoxynucleotides (ASON) on the growth of HL-60 cells in nude mice].

OBJECTIVE: To study the inhibition effect of cyclin G(1) antisense oligodeoxynucleotides (ASON) on the growth of HL-60 cells in nude mice. METHODS: (1) Nude mice were divided into control group, sense oligodeoxynucleotides (SON) group and ASON group. After (60)Co radiation, with HL-60 cells SON group and ASON group were subcutaneously innoculated; (2) The weight and volume of tumors were continually measured; (3) The morphology of tumor cells was observed by microscope; (4) The protein and mRNA expression levels of cyclin G(1) were determined by flow cytometry (FCM) and reverse transcription polymerase chain reaction (RT-PCR); (5) The cell apoptosis was detected by electron microscopy and FCM. RESULTS: (1) The inhibition rate of tumor in ASON group was 69.4%. In ASON group, the wight and volume of tumor were significantly lower than those in SON group and control group. (2) The HL-60 cells in ASON group showed morphologically smaller nuclei, less mitosis, less heteromorphosis and apoptosis. CONCLUSION: The cyclin G(1) ASON can inhibit the growth of HL-60 cells in nude mice and induce apoptosis.

[Effect of liposomal transfection of cyclin A antisense oligodeoxynucleotide (ASON) on HL-60 cell proliferation and apoptosis].

OBJECTIVE: To explore the effect of liposomal transfection of cyclin A antisense oligodeoxynucleotide (ASON) on HL-60 cell proliferation and apoptosis. METHODS: By liposomal transfection, cyclin A ASON was co-cultured with HL-60 cells, the cell growth curve was determined by MTT assay and cell apoptosis electron-microscopy in situ cell apoptosis detection kit (POD), the protein and mRNA of cyclin A and bcl-2 were measured by FACS and RT-PCR, the role of cyclin A ASON in the development of leukemia was tested by the tumor formation in nude mice. RESULTS: (1) In the cyclin A ASON liposomal transfection group (group A), the proliferation of HL-60 cell was significantly inhibited as compared to those in cyclin A ASON group (group B) (68.9% vs 24.8%) (P < 0.01). (2) The expressions of cyclin A and bcl-2 of group A were significantly lower than those in the control group (1.1% vs 38.8%, P < 0.01; 21.9% vs 65.0%, P < 0.01, respectively), and the DNA ladder and apoptosis body was displayed. (3) In group A, the rate of tumor formation in nude mice was lower, the time for tumor formation was longer and the volume of tumor was smaller than those in control group. CONCLUSION: Liposomal transfection of cyclin A ASON can inhibit in vitro proliferation of leukemia cells and induce in vivo apoptosis of the tumor cell, which might provide a new target for gene therapy.