ABSTRACT
Objective:To investigate the expression of Galectin-7 in the bronchial mucosa of asthmatic children and its effect on the apoptosis of bronchial epithelial cells.Methods:Bronchial mucosa of asthmatic children and children with bronchial dilation were collected and the expression of Galectin-7 was detected by Western blot.Human bronchial epithelial cells were cultured in vitro,the cells were transfected with Galectin-7 siRNA to interfere the Galectin-7 expression,while siRNA control was transfected as the control group.The experiment was divided into normal group,control group,infected group and experimental group.The normal group was normal human bronchial epithelial cells,the cells in the control group was transfected with siRNA control,the infected group was infected with RSV,the experimental group was transfected with Galectin-7 siRNA and infected with RSV.After 24h culture,Galectin-7 protein expression and cell apoptosis were detected in the cells of each group.Western blot was used to detected the expression of Bcl-2,Bax,STAT3 and p-STAT3.Results:The expression of Galectin-7 in bronchial mucosa of asthmatic children was significantly higher than that of the non asthmatic children (P＜0.01).There was no significant difference in the Galectin-7 level between the normal group and the control group (P＞0.05).The levels of Galectin-7,Bax and apoptosis in the infected group were significantly higher than those in the normal group,while the levels ofp-STAT3 and Bcl-2 were significantly lower than those in the normal group (P＜0.01).The levels of Galectin-7,Bax and apoptosis in the experimental group were significantly lower than those in the infected group,while the levels of p-STAT3 and Bcl-2 were significantly higher than those in the infected group (P＜0.01).Conelusions:The expression of Galectin-7 was up-regulated in the bronchial mucosa of asthmatic children,which might promote the apoptosis of bronchial epithelial cells by activating STAT3.
ABSTRACT
Objective To prepare the mouse anti recombinant human Galectin-7 antibody and the antibody was characterized in bladder cancer.Methods The gene coding for Galectin-7 was amplified by PCR from the cDNA of human foreskin cells and cloned into prokaryotic expression vector pET28a.Then the recombinant plasmid pET28a/Galectin-7 was transformed into E.coli BL21 (DE3)and expressed under IPTG induction.The recombinant Galectin-7 was purified through Ni2+-NT agarosegel column and the purified Galectin-7 used as imunogen to imunize the mouse.The titer and specificity of the anti-Galectin-7antibody from the mouse were analyzed by ELISA,Western blot and immunohistochemistry,respectively.Results The recombinant Galectin-7 was successfully expressed and purified,and the polyclonal ani-Galectin-7 antibody was suc-cessfully prepared.The titer of the antiserum was 1∶32 000 by ELISA.Western blot analysis showed this antiboday reacted specifically with Galectin-7.Immunohistochemistry analysis showed the antibody could recognize the native Galectin-7 in the human bladder cancer tissue.Conclusion The preparation recombinant Galectin-7 protein was as immunogen in rabbits.It was successful to produce high titer and high specificity of anti Galectin-7 polycolonal antibody.
ABSTRACT
Purpose To study the expression of Galectin-7 in squamous carcinoma of the cervix and its correlation with HPV 16 infec-tion in Uyghur and Han women. Methods The expression of Galectin-7 in paraffin-embedded cervical tissues collected from Uyghur women including 21 patients with cervicitis and 49 patients with squamous carcinoma of the cervix and Han women including 25 patients with cervicitis and 29 patients with squamous cervical cancer was evaluated by immunohistochemical SP method. The infection of HPV 16 was evaluated by PCR amplification of HPV DNA from the corresponding cervical tissue samples. The expression of levels of Galec-tin-7 in cervical tissues was analyzed according to the corresponding integrated optical density ( IOD) value of Galectin-7 in cervical le-sions with professional image analysis software Image-Pro Plus (IPP) 6. 0. Results PCR detection of HPV 16 in Uyghur patients from Kashghar showed that 20 patients were positive and 29 cases were negative for HPV 16 in squamous cervical cancer patients whereas in patients with cervicitis, only 5 cases were positive, 16 cases were negative for HPV 16. Furthermore the detection of HPV 16 in Han patients showed that 2 cases were positive and 27 cases was negative in patients with squamous cervical cancer patients, and all the ca-ses were negative for HPV 16 in patients with cervicitis. The expression of Galectin-7 in squamous cervical cancer tissues was signifi-cantly lower than that in the cervicitis patients (P<0. 05). The expression of Galectin-7 in HPV 16 positive squamous cervical cancer patients was significantly lower than that in HPV 16 negative patients ( P<0. 05 ) . There was no obvious correlation between the ex-pression levels of Galectin-7 in squamous cervical cancer patients with the differentiation degree and disease oneset age both in Uyghur and Han patients. Conclusion Compared with cervicitis tissues, Galectin-7 expression in squamous cervical cancer is down-regulated in Uyghur women from Kashghar region implying that the expression of Galectin-7 may play a certain role in the occurrence and develop-ment of squamous cervical cancer, and its detection has a certain reference value for the diagnosis of cervical cancer for Uyghur women.