ABSTRACT
Exogenous wild-type p53 (wt-p53) tumor suppression increases the sensitivity of tumor cells to radiotherapy and chemotherapy. An iodized oil emulsion was used as a p53 vector for intra-arterial gene delivery to treat hepatic tumors. Whether the chemotherapeutic agent or the iodized oil affects exogenous wt-p53 activity remains poorly understood. In the present study, the early therapeutic response of rAd/p53, combined with 5-fluorouracil (5-FU) or with iodized oil, was observed in a human colon cancer model. Allograft models in 82 nude mice with human colon carcinoma SW480 were divided randomly into four groups and administered with physiologic saline, rAd/p53, rAd/p53+5-FU, and rAd/p53+iodized oil by intratumoral injection. At 24, 48, 72, 120, and 168 h after treatment, p53 expression, the Ki-67 index (KI), and the degree of tumor necrosis were assessed. The p53 expression and tumor necrosis in the therapeutic groups were higher than those in the control group. p53 expression reached its peak at 120 h in the rAd/p53 group, at 72 h in the rAd/p53+5-FU group, and at 48 h in the rAd/p53+iodized oil group. The p53 expression in the rAd/P53+5-FU group and the iodized oil group was significantly higher than those in the rAd/P53 group at 24 and 48 h. The results revealed that tumor necrosis is positively correlated with p53 expression. The KI of the rAd/p53+5-FU group increased significantly at 24 h. 5-FU and iodized oil increase the anticancer effect of rAd/p53, and 5-FU combined with rAd/p53 has a synergistic anticancer effect.
Subject(s)
Antimetabolites, Antineoplastic/administration & dosage , Colonic Neoplasms/genetics , Colonic Neoplasms/therapy , Fluorouracil/administration & dosage , Genetic Therapy , Iodized Oil/administration & dosage , Tumor Suppressor Protein p53/genetics , Animals , Humans , Mice , Mice, Nude , Transplantation, HeterologousABSTRACT
This study examined the expression of human leukocyte antigen (HLA)-G and HLA-I (which includes HLA-A, -B, -C, -E and -F, but is without HLA-G) in the cleavage embryo and its supernatant, and related the results to embryo development including growth rate and grade. In total, 136 day-3 cleavage embryos were used for detection of HLA-G and 24 embryos for HLA-I without HLA-G by immunohistochemistry. The expression of HLA-I was examined by western blot in the lysates of a further 63 day-3 cleavage embryos; soluble HLA-I in the culture supernatant of embryos with detectable HLA-I was also examined by western blot. It was found that 90 of 136 (66.2%) cleavage embryos expressed HLA-G, whereas 23 of 24 (95.8%) embryos expressed HLA-I without HLA-G. HLA-G expression typically showed an even and symmetrical pattern of distribution in each blastomere. HLA-I without HLA-G in cleavage-stage embryos is typically scattered around the blastomere surface. The expression of HLA-G but without HLA-I in cleavage-stage embryos was significantly associated with embryo grade (P < 0.001) and cell number (P = 0.03). In conclusion, HLA-I is expressed on day-3 cleavage embryos, and HLA-G expression on preimplantation embryos is related to embryo development, including embryo growth rate and embryo grade.
Subject(s)
Cleavage Stage, Ovum/metabolism , Embryo, Mammalian/metabolism , Histocompatibility Antigens Class I/genetics , Blastocyst/cytology , Blastocyst/metabolism , Cell Count , Cells, Cultured , Culture Media/chemistry , Culture Media/metabolism , Embryo, Mammalian/cytology , Embryonic Development/genetics , Gene Expression Regulation, Developmental , HLA Antigens/genetics , HLA Antigens/metabolism , HLA-G Antigens , Histocompatibility Antigens Class I/analysis , Histocompatibility Antigens Class I/metabolism , Humans , Quality Control , Time FactorsABSTRACT
OBJECTIVE: To explore the association of altered expression of annexin IV in human endometrium during the implantation window and endometrial receptivity. METHODS: A comparative proteomic strategy, in a combination of two-dimensional fluorescence difference gel electrophoresis (2D-DIGE) and matrix-assisted laser desorption/ionization time of flight mass spectrometry (MALDI-TOF-MS), was adopted to search for proteome alternations of pre-receptive (day LH + 2) versus receptive (LH + 7) endometria. The location and abundance of the identified differentially expressed protein- annexin IV were analyzed by immunostaining and western blot. RESULTS: By comparing protein profiles of LH + 2 and LH + 7 samples, we found a protein up-regulated 2.12 times in LH + 7 samples, with a relative molecular weight of 36,000 and an isoelectric point near pH 5.8. It was characterized using mass spectrometry and was identified as annexin IV. Immunohistochemical analysis revealed an altered localization of annexin IV--in the epithelia on day LH + 2, and both in the epithelia and stroma cells on day LH + 7. Protein levels of annexin IV were up-regulated on day LH + 7 compared with that on day LH + 2 by Western blot. Integrated optical density of the object (OPTDI) was 46.249 +/- 32.376 and 249.507 +/- 31.959, respectively (P = 0.004). CONCLUSIONS: Our study indicates endometrial samples obtained by microbiopsy are available for proteomics studies. It seems possible that the increased expression of annexin IV during the implantation window plays an important role in the morphological differentiation of the uterus to the receptive state.
Subject(s)
Annexin A4/biosynthesis , Embryo Implantation , Endometrium/metabolism , Menstrual Cycle/physiology , Adult , Blotting, Western , Electrophoresis, Gel, Two-Dimensional/methods , Embryo Implantation/physiology , Female , Humans , Immunohistochemistry , Pregnancy , Proteomics , Receptors, Estrogen/metabolism , Receptors, Lymphocyte Homing/metabolism , Receptors, Progesterone/metabolism , Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization/methods , Time FactorsABSTRACT
OBJECTIVE: To study expression of insulin receptor substrate 1 (IRS-1) and phosphorylation of tyrosine in adipose tissue of polycystic ovary syndrome (PCOS) to approach the mechanism of insulin resistance (IR) in PCOS at tissue and cellular level. METHODS: IRS-1 expression and tyrosine phosphorylation of adipose tissue were studied with immunoprecipitation, Western-blot and ECL immunoblotting. RESULTS: The differences of IRS-1 expression in adipose tissue among the obese PCOS group (82 +/- 15)%, the non-obese PCOS group (79 +/- 18)%, the obese control group (75 +/- 19)% and the non obese control group (70 +/- 19)% were not significant (P > 0.05). IRS-1 tyrosine phosphorylation in adipose tissue in the obese PCOS group [(52 +/- 23)%, P < 0.001], the obese control group [(45 +/- 22)%, P < 0.01] and non-obese PCOS group [(70 +/- 25)%, P < 0.05] were markedly lower than that in the non-obese control group (88 +/- 12)%. The obese control group had more reduction than non-obese control group (P < 0.05). The differences between the obese PCOS group and the obese control group or the two PCOS groups were not significant (P > 0.05). CONCLUSION: IRS-1 tyrosine phosphorylation in adipose tissue in the two PCOS groups decreased markedly than in the non-obese control group. It might be involved in the disorder of insulin-signaling transduction downstream of insulin receptor and the occurrence of IR.
