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Objective To analyze the mutation status of K‐ras gene in the patients with gastric cancer to provide the guidance for the personalized therapy of gastric cancer .Methods The nested and Cold‐PCR were adopted to analyze the K‐ras gene mutation status in 314 cases of gastric cancer .Results In 314 cases of gastric cancer ,the total mutation rate of K‐ras gene was 7 .32% .The mutation rate was 0% in 19 plasma samples and 7 .80% in 295 tissue samples .The types of mutation included G12D ,G13D ,G12V , the mutation rate of K‐ras gene had no statistically significant difference between the two kinds of different samples (P=0 .206 1);the mutation rate was 6 .79% in 221 male patients ,the types of mutation included G12D ,G13D ,the mutation frequency is 8 .60% in 221 female patients ,the types of mutation include G12D ,G13D ,G12V ,the mutation rate of K‐ras gene had no statistically signifi‐cant difference between different genders (P=0 .573 1);the mutation rate was 6 .67% in 45 youth patients ,the types of mutation included G12D ,G13D ,the mutation rate was 7 .87% in 127 middle age patients ,the types of mutation included G12D ,G13D ,G12V , the mutation rate was 7 .04% in 142 old age patients ,the types of mutation included G12D ,G13D ,there was no statistically signifi‐cant difference among different age patients (P=0 .995 3) .Conclusion The mutation rate of K‐ras gene is 7 .32% in 314 cases of gastric cancer ,the main mutation types include G12D and G13D ,and the mutation rate of K‐ras gene has no significant difference a‐mong different samples ,between different sexes and among different ages .
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Objective To analyze the polymorphism of excision repair cross-completion gene 1(ERCC1) in colorectal cancer pa-tients ,and to provide guidance for personalized therapy of colorectal cancer .Methods The polymorphism of ERCC1 in 259 patients with colorectal cancer was detected by PCR .Results The genotype of ERCC1 was mainly C/C(55 .71% ) in male patients ,and then C/T(37 .86% ) ,the genotype of T/T accounted for 6 .43% .The genotype of ERCC1 was also mainly C/C(59 .66% ) in female pa-tients ,and then C/T(36 .97% ) ,the genotype of T/T accounted for 3 .36% .The genotype was mainly C/C(51 .72% ) aged 25 to 44 patients ,and then C/T(41 .38% ) ,the genotype of T/T accounted for 6 .90% .The genotype was mainly C/C(57 .95% ) aged more than 44 to 59 patients ,and then C/T (40 .91% ) ,the genotype of T/T accounted for 1 .14% .The genotype was mainly C/C (58 .45% ) aged more than 59 patients ,and then C/T(34 .51% ) ,the genotype of T/T accounted for 7 .04% .Conclusion The poly-morphism of ERCC1 is mainly C/C in colorectal cancer patients .
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The use of rnicroarrays of oligonucleotides or cDNA is considered to be a promising approach for DNA and RNA sequence analysis, diagnostics of genetic diseases, gene polymorphism studies and analysis of gene expression. To manufacture cDNA microarrays the samples were printed onto glass microscope slides treated with poly-L-lysine, and then the slides were processed by heat and UV light treatment to attach the cDNA sequence to the glass surface. But the immobilization efficiency of cDNA on the glass surface was low. A simple procedure for manufacture cDNA microarrays on a slide treated with 3-aminopropyltrimethoxysilane is described. The efficiency for attaching cDNA to the amino-modified slides is greater than that to the slides treated with poly-L-lysine. The cDNA microarray made by the amino-modified slides is stable for use in 80℃, 75 % humidity, 3 600Lx light, exposure in air, respectively.
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Background and purpose:Wuxing soup is popular for it's anti-cancer effect in folk medicine and deserved to be further studied by modern scientific methods.This research aimed to explore its anti-cancer effect and to study the influence on the immunity of melanoma in mice. Methods :Inhibition of different ingredients and concentrations of Wuxing soup on the growth of the mouse melanoma B16 cells was detected by MTT in vitro.Animal experiment was performed to determine its anti-cancer effect in vivo.C57/BL6 mice were randomly divided into three groups after vaccination of B16 cell,and then given intragastrically with different soups for 25 days.All mice were killed on day 26 after inoculation.The weight of tumors were recorded.The anti-tumor immune function was measured by T lymphocyte transforming assay and NK killing assay.The different effect of different ingredients was also observed. Results :Our result showed that different ingredients soup exerted different inhibition on B16 cell growth and Wuxing soup was the strongest one of all and in dose-dependent manner in vitro.The animal experiment indicated that different ingredients soup has different inhibition on melanoma,the soup-treated mouse display improved T lymphocyte transforming assay and NK killing ability.Among the different soups,Wuxing soup showed the strongest anti-cancer effect and immune enhancement. Conclusions :Wuxing soup is an effective anti-cancer agent in melanoma mice and can enhance the immunity of the mice with melanoma.
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Purpose To introduce a method to isolate cDNA clones using bacteriophage P1-derived artificial chromosome (PAC) or bacterial artificial chromosome (BAC) as probe for hybridization and try to find some novel genes related to hepatocellular carcinoma. Methods PAC 579 (D17S926 locus) and BAC 1529 (D17S1529 locus) in the deletion region of chromosome 17p13.3 in human hepatocellular carcinoma were chosen to screening the human liver cDNA library as probe for hybridization. The isolated positive cDNA clones were partially sequenced, then analyzed by computer comparison and Southern blot. Results After three cycles of screening, 78 and 8 candidate positive cDNA clones were isolated using PAC 579 and BAC 1529 probes respectively. Further analysis indicated 18 cDNA clones isolated by PAC 579 probe and 5 cDNA isolated by BAC 1529 probe were potential novel genes related to hepatocellular carcinoma. Conclusions The isolation of cDNA clones using PAC and BAC probes is effective and practical.
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Background and purpose:Hepatocellular carcinoma suppressor gene-1(HCCS1)is a potential hepatocellular carcinoma supressor gene,and it also plays an important role in sorting of some cytoplasmic proteins.Its carcinoma suppressor function may be related to its sorting function.So it is important to identify the minimum region of functional sequence in HCCS1 that related to the transportation.Methods:The expression vectors containing various lengths of HCCS1 gene were constructed and transfected into HeLa cells mediated by liposomes.The localizations of different HCCS1 fragments and the colocalizations of M6PR with various truncated HCCS1 were determined by laser scanning confocal microscopy,respectively.Results:10 different expression vectors containing various lengths of HCCS1 gene were successfully constructed,it had been shown that the polar localization of HCCS1 protein and the co-localizations with M6PR disappeared when HCCS1 gene was truncated to 1 120 bp from the 3'end by laser scanning confocal microscopy.Conclusions:We identified the minimum localization of the HCCS1 functional sequence that related to the transportation.