| Objective:To explore the expression of autophagy related protein p62?Beclin-1 and LC-3 in cervical tissues,may be suggested the molecular biology sign in the development of cervical cancer,also provide the direction for targeted therapy in cervical cancer.Methods:1.Choose the persons who treated by operation or examined under colposcope,in the Second Affiliated Hospital of Shanxi Medical University,from January 2015 to January 2017,select 60 cases which have been diagnosed as cervical squamous cell carcinoma as case group,30 cases which have been diagnosed as chronic cervicitis as control group;2.select the above patients' s pathological section in the Department of Pathology,using immunohistochemical method,qualitative detection the expression of p62 ? beclin-1 and LC-3;3.select the above patients' s organization in the liquid nitrogen,using protein lysis to extract the desired protein,Western blot method was used to quantitatively observe the expression of p62?Beclin-1 and LC-3;4.collect the above patients' s relevant information about the age,clinical pathological stage,pelvic lymph node metastasis,tumor size,differentiation and so on,to analyze if there were some correlation with p62?Beclin-1 and LC-3;5.use SPSS18.0 statistical analysis software for data analysis: the relative expression of p62?bclin-1 LC3 were expressed by mean±SEM;in the experimental group and the control group,the analyzation of difference expression in the three indicators by chi square test;the analysis of the three indicators' s correlation using Spearman test;the difference expression of P62?Beclin-1 and LC-3with the clinical characteristics in cervical cancer,analyzing by chi square test;the consistency analysis in the positive rate and negative rate of expression in p62?Beclin-1and LC-3 using the kappa test in immunohistochemical results;Test level ?=0.05.Results:1.Immunohistochemical results: the autophagy marker protein p62 expressed mainly in the cytoplasm,the positive expression rate in each group are as follows:control group 36.7%(11/30),cervical cancer 68.3%(41/60),the result is meaningful(P < 0.05);The expression of Beclin-1 have been detected in cytoplasm and cell membrane,but mainly expressed in the cell membrane,the positive rates in control group 76.7%(23/30),cervical cancer 28.3%(17/60),the result is meaningful(P < 0.05);the expression of LC-3 mainly in cytoplasm and nucleus,the expression in cell membrane was very small,the positive expression rate in control group 73.3%(22/30),cervical cancer 35.0%(21/60),the result is meaningful(P < 0.05).2.Western blot showed that the relative expression levels of p62 in the two groups were: control group(0.289±0.021),cervical cancer(0.563±0.031);The relative expression of Beclin-1 for control group(0.547±0.063),cervical cancer(0.310±0.018);the relative expression of LC-3 for control group(0.679±0.063)?cervical cancer(0.303±0.024).The result is meaningful(P < 0.05).3.The difference expression of P62 ? Beclin-1 and LC-3 with the clinical characteristics: the positive expression of P62 in the group of tumor's diameter:>4cm64.3%(18/28)? ?4cm 65.6%(21/32),the result is meaningful(P < 0.05);in high differentiation,moderate differentiation and low differentiation,the positive expression rate were 57.1%(15/28),94.7%(18/19),84.6%(11/13),the result is meaningful(P <0.05).The positive expression of Beclin-1 in pelvic lymph node metastasis rate:yes 9.09%(1/11),no 44.8%(22/49),the result is meaningful(P < 0.05);in high differentiation,moderate differentiation and low differentiation,the positive expression wererespectively: 60.7%(17/28),26.3%(5/19),0%(0/13),the result is meaningful(P < 0.05).The positive expression of LC-3 in the group of tumor's diameter:>4cm 60.7%(17/28),? ?4cm 71.9%(23/32),the result is meaningful(P < 0.05);the positive expression in pelvic lymph node metastasis rate:yes 18.2%(2/11),no 42.9%(21/49),the result is meaningful(P < 0.05);in high differentiation,moderate differentiation and low differentiation,the positive expression were respectively: 60.7%(17/28),15.8%(3/19),15.4%(2/13),the result is meaningful(P < 0.05);The positive expression rates in different clinical stages: I-IIa 47.6%(20/42)? IIB-IV 16.7%(3/18),the result is meaningful(P < 0.05).4.Spearman was used for the analysis of correction with P62 Beclin-1 LC-3: The expression of p62 was not correlated with Beclin-1 and LC-3(r=-0.312 p=-0.071;r=-0.276 p=0.067);the expression of Beclin-1 was positively correlated with LC-3(r=0.443 P < 0.05)In the immunohistochemical group,Kappa test was used to analyze the consistently expression rates of Beclin-1 and LC-3,kappa= 0.548,indicating that the expression of the two protein was higher consistent in different tissues.Conclusion:1.In immunohistochemistry and Western blot,the expression of P62 was significantly up-regulated in the cervical cancer group,on the contrary,Beclin-1 and LC-3 was down-regulated in the cervical cancer group;suggest that from the quantitative or qualitative level,the autophagy activity has been reduced in cervical cancer.2.From the analysis of relationship between p62?Beclin-1 and LC-3 with clinical manifestations of cervical cancer: the expression of p62 has the relationship with the tumor size(?2 =6.323 p=0.031)and cervical carcinoma(?2 =6.532 p=0.033);Beclin-1with pelvic lymph node metastasis(?2 =8.417 p=0.007)and the degree of differentiation the tumor tissue(?2 =9.734 p=0.000);LC-3 was correlated with tumor size(?2 =7.441p=0.040),pelvic lymph node metastasis(?2 =5.093 p=0.037),the degree of tumor differentiation(?2 =13.001 p=0.001)and the clinical stage of cervical cancer(?2=5.003 p=0.020);This indicates that the change of autophagy activity is also involved inthe development of cervical cancer.3.Through the correlation analysis of the three indicators: The expression of p62 and Beclin-1 and LC-3 had no significant correlation(r=-0.312 p=-0.071;r=-0.276 p=0.067);the expression of Beclin-1 and LC-3 was positively correlated(r= 0.443,P < 0.05).The Beclin-1 and LC-3 has high consistency in reflecting the different tissues the expression of protein. |
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