Expression of galectin-3 in human breast cancer tissues and its effect on malignant biological behavior of MCF-7 cells / 中国肿瘤生物治疗杂志

@# Objective: To investigate the expression of galectin-3 protein in human breast cancer tissues and the effect of silencing galectin-3 gene on the migration, invasion and apoptosis of human breast cancer MCF-7 cells. Methods: The relative expression of galectin-3 protein in 15 cases of breast cancer tissues and corresponding para-cancerous tissues were detected by Western blotting; The expression of galectin-3 protein in paraffin sections of 100 cases of breast cancer tissues were detected by immunohistochemistry, and the correlation between galectin-3 expression and the clinicopathological characteristics of breast cancer patients was also analyzed. Galectin-3 siRNA were transfected into human breast cancer MCF-7 cells by liposome, then Real-time PCR and Western blotting were used to detect the mRNA and protein expression of galectin-3. The effect of galectin-3 gene silencing on cell migration and invasion ability of MCF-7 cells were detected by Transwell method. The effect of galectin-3 gene silencing on apoptosis of MCF-7 cells were detected by flow cytometry. Results: Western blotting detection showed that the relative expression of galectin-3 protein in breast cancer tissues were significantly higher than that in para-cancerous tissues (P<0.05); Immunohistochemistry detection showed that the positive expression rate of galectin-3 protein in breast cancer tissues was 67.00%, the positive expression rates in the lymph node metastasis, hormone receptor (ER, PR) negative groups were significantly higher (P<0.05), and the positive expression rate of galectin-3 protein were increased with the increase of TNM stage and histological grade (P<0.05); Galectin-3 siRNA transfection could significantly reduce the mRNAand protein expression of galectin-3 in MCF-7 cells (P<0.05), and reduce the invasion and migration ability but significantly improve the rate of apoptosis of MCF-7 cells (P<0.05). Conclusion: Galectin-3 is highly expressed in breast cancer tissues, and its silence can inhibit the invasion and metastasis of MCF-7 cells and induce apoptosis of MCF-7 cells. Galectin-3 can be used as a new target for biological therapy of breast cancer.

HSF1 inhibits heat stress-induced apoptosis in Raw264.7 macrophages / 中南大学学报(医学版)

OBJECTIVE@#To observe the effect of heat shock factor 1 (HSF1) on heat stress-induced apoptosis in Raw264.7 macrophages.@*METHODS@#Raw264.7 cells transfected with pcDNA3.1 and pcDNA3.1-HSF1 were exposed to heat stress (42.5 degrees C +/- 0.5 degrees C) for 1 h and recovered at 37 degrees C for 6, 9, 12, and 24 h respectively. Flow cytometry (FCM), Hoechst 33258 staining and DNA ladder assays were performed to assess the apoptosis.@*RESULTS@#After heat stress, FCM showed that apoptotic cells were increased significantly and reached the peak at 9 h in Raw 264.7 cells transfected with pcDNA3.1, and were characterized with classical morphologic changes including apoptotic body and nuclear condensation. Agarose gel electrophoresis showed that "DNA ladder" could be observed clearly at 6, 9, and 12 h after the heat stress. But the overexpression of HSF1 could reduce the number of apoptotic cells and inhibit DNA fragmentation.@*CONCLUSION@#HSF1 can inhibit heat stress-induced apoptosis in Raw264.7 macrophages.

Screen of inflammatory genes regulated by heat shock factor 1 and corroboration with SOCS3 gene / 中南大学学报(医学版)

OBJECTIVE@#To screen the inflammatory mediators genes regulated by HSF1, and explore the mechanism of downstream genes regulated by HSF1.@*METHODS@#HSF-/- and HSF1+/+ mice were injected with 15 mg/kg LPS intraperitoneally (ip), respectively, and were treated as previous after HSR. The total RNA of lung tissues were extracted and filtrated by SuperArray gene Microarry. The promoter of candidate genes were analyzed by transcription element search software to search for heat shock element (HSE). Select the suppressor of cytokine signaling 3 (SOCS3) with HSE. Macrophage cells were stimulated with 400 ng/mL LPS, and were treated as previous after HSR, then the total RNA was extracted respectively. RT-PCR and northern blot assay were performed to detect the expression levels of SOCS3 mRNA.@*RESULTS@#Fifteen genes were repressed by HSF1, including 9 genes with complete HSE. Eleven genes were accelerated by HSF1 possibly, including 8 genes with complete HSE. The promoter of SOCS3 gene contained one complete HSE. LPS stimulation obviously increased the levels of SOCS3 mRNA in macrophages of RAW264.7 mice, which was inhibited by HSR and over-expression of HSF1.@*CONCLUSION@#HSR or HSF1 inhibits LPS induced expression of SOCS3 mRNA; HSF1 might inhibit LPS-induced expression of SOCS3 mRNA by binding to HSE in the promoter of SOCS3 gene.