Relationship Between GLI3 Protein Expression And Clinicopathological Features In NPC Tissues

Objective: To explore the relationship between GLI 3 protein expression in the cytoplasm and nucleus of NPC tissue,the level of different forms of GLI 3 protein(i.e.full-length GLI 3 protein,truncated GLI 3 R)in NPC tissue and clinicopathological features of NPC patients.Methods: First of all,the study subjects were 101 patients admitted to Nanxishan Hospital(Nanxishan Hospital affiliated to Guilin Medical University)from June 2013 to June 2019.In addition,the nasopharyngeal biopsy mucosal tissues of 20 patients pathological diagnosed with mild chronic nasopharyngeal mucositis epithelium by the hospital at the same time were selected.The above biopsy tissues have been made as paraffin-embedded tissue blocks and archived in the Department of Pathology department of the hospital.Patient medical records and diagnosis and treatment information are perfected.GLI3 protein expression levels in these tissues were determined by immunohistochemistry.Based on the intensity of immune response staining and the percentage of positive cells,the expression level of NPC and Immuno-Reactive-Score System(IRS)was used to compare the relationship between GLI3 in NPC nucleus and cytoplasm and the clinicopathological characteristics of tumors.Secondly,western blot(WB)technology was used to detect the expression of unprocessed full-length GLI 3 protein(GLI3 full-long,GLI3FL),processed GLI 3 R(GLI3 repressor,GLI 3 R)in 27 NPC tissues,In addition,7 patients with mild chronic nasopharyngeal mucositis diagnosed by pathology in the hospital during the same period were selected for nasopharyngeal biopsy of mucosal tissue.Analyze the relationship between the expression levels of GLI3 FL and GLI3 R proteins in epithelial tissue between nasopharyngeal carcinoma and mild chronic nasopharyngeal mucositis,as well as the relationship between the expression of GLI3 FL and GLI3 R in nasopharyngeal carcinoma tissue and clinical pathological characteristics.Results:(1)The expression level of GLI3 protein in the cytoplasm and nucleus of nasopharyngeal carcinoma tissue was higher than that in the epithelial tissue of mild chronic nasopharyngeal mucositis(cytoplasm:6.28 ± 3.21 V.s 2.73 ± 2.02;nucleus: 6.23 ± 2.82 V.s 1.93 ±1.28;both P<0 01)。 22(21%)cases had high expression of GLI3 protein in the nucleus of nasopharyngeal carcinoma,while 79(78%)cases had low expression;31(31%)cases had high expression of GLI3 protein in the cytoplasm,while 70(69%)cases had low expression;20(100%)cases of mild chronic nasopharyngeal mucositis had low expression of GLI3 protein in epithelial cells,while high expression of GLI3 protein in cytoplasm accounted for 2(10%)cases and low expression of GLI3 protein in cytoplasm accounted for 18(90%)cases;The expression level of GLI3 protein in the cytoplasm of nasopharyngeal carcinoma tissue was statistically significant compared with the depth of tumor invasion(P<0.01).(2)Using WB detected GLI3 FL and GLI3 R proteins in 27 cases of nasopharyngeal carcinoma tissue(GLI3FL: 0.7809 ± 0.1441,GLI3R: 0.5891 ± 0.1125,GLI3R/GLI3FL: 0.7597 ± 0.0791,GLI3FL/GLI3R: 1.3229 ± 0.1354,respectively)and 7 cases of mild chronic nasopharyngeal mucosal inflammation epithelial tissue(GLI3R:0.2453 ± 0.0614,GLI3R/GLI3FL: 0.8881 ± 0.1764,GLI3FL/GLI3R:1.1584 ± 0.1924,respectively).The expression levels of GLI3 FL and GLI 3 R protein(both P <0.01)and GLI3 FL / GLI 3 R ratio(P <0.05)were significantly higher in NPC tissues than those of mild chronic nasopharyngeal mucositis.There was no statistically significant difference in GLI3R/GLI3 FL ratio(P=0.105)between above two group.The detection results of GLI3 expression in 27 nasopharyngeal carcinoma tissues by WB showed that the expression levels(?χ± S)of GLI3 R protein in TNM stage II+III and IV of nasopharyngeal carcinoma were0.5804 ± 0.1086 and 0.6017 ± 0.1222,respectively(P=0.639),and the expression levels in T1+T2 and T3+T4 were 0.5036 ± 0.1387 and0.6394 ± 0.0501,respectively(P=0.013).No significant changes in GLI3 FL levels were observed as nasopharyngeal carcinoma progressed.The GLI3R/GLI3 FL ratio(?χ± S)was 0.7254 ± 0.0619 and 0.8096± 0.0769 in tumors II+III and IV,respectively(P=0.004);The GLI3FL/GLI3 R ratio(?χ± S)was 1.3753 ± 0.1131 and 1.2466 ±0.1329 in II+III and IV tumors,respectively(P=0.012).There was a significant correlation between GLI3 FL and GLI3 R protein expression levels(r=0.645,p=0.0005).Conclusion: The expression of GLI3 protein was up-regulated in the nucleus and cytoplasm of nasopharyngeal carcinoma.The expression levels of GLI3 FL and GLI 3 R protein and the GLI3 FL / GLI 3 R ratio were significantly higher in NPC tissues than in mild chronic nasopharyngeal mucositis epthilial tissues.The GLI3FL/GLI3 R ratio in nasopharyngeal carcinoma tissue is related to clinical staging of the tumor,and the expression level of GLI3 protein in the cytoplasm of cancer cells is related to the degree of tumor infiltration.This suggests that the different forms of GLI3 protein exist,especially the GLI 3 R content as well as ratio of GLI3FL/GLI3 R,which may play an important role in the invasion and metastasis of nasopharyngeal tumors.