| Objectives:The associations between of the expressions of calcium-activated chloride channel TMEM16A and the development,prognosis and clinic pathological characteristics for several common malignant epithelium tumors and colorectal adenomas were explored.Aim to illustrate 1)the intervention effect of the traditional Chinese medicine:Jianpi-Huayu decoction on the recurrence of colorectal adenomas after operation and regulation of the expression of TMEM16A;2)the role of TMEM16A,HER2 and EGFR expressions during the development and prognosis of colorectal adenocarcinoma;3)the function and underlying molecular mechanisms of TMEM16A expression during the development and prognosis of Non-small cell lung cancers.Methods:1.From November 2011 to October 2014,60 patients,performed endoscopic resection of colorectal adenoma,and confirmed as colorectal adenoma with intraepithelial neoplasia by pathology diagnosis,were collected sequentially.The 60 patients were randomly assigned into two groups:(1)Jianpi-Huayu decoction(30 cases)were applied to the treatment group after one week of the endoscopy resection;(2)the other 30 cases were used as placebo group without the decoction treatment.After one year,we examined the tumor number,size,recurrence,and observed the pathological morphology of tumor cell under the microscope,and semi-quantitated the expression of TMEM16A,COX-2,Ki67 proteins by using IHC(immunohistochemistry).2.From March 2016 to March 2017,semi-quantitated the expression of TMEM16A,Her-2 and EGFR proteins by using IHC from the Paraffin embedded tissues of 157 surgical resection of colorectal cancer and 30 normal colorectal mucosa.At the first author/time(in China),systematically analyzed the associations of the expressions of TMEM16A,HER-2 and EGFR with the development,prognostic and clinic pathological determinants of colorectal cancers.3.From September 2011 to March 2015,83 paraffin embedded specimens from surgical recession of Non-small cell lung cancers and adjacent tissues were collected to:(1)Semi-quantitate the expression of TMEM16A and EGFR proteins by using IHC;(2)study the expression of TMEM16A proteins by using western blotting;(3)detect the amplification by using real-time fluorescence quantitative PCR.Additionally,following up were performed for these 83 patients.Results:1.There were significant difference(P<0.05)for index of both the number and the size of colorectal adenoma between the treatment group(applied Jianpi-Huayu decoction)and the placebo group.The number and size of tumors at treatment group were less than those for placebo group(t value were 4.980,4.788 respectively,P<0.01).2.The adenoma recurrence rates were 16%(4/25)and 57.1%(16/28)respectively for treatment and placebo group,there was a significant difference(P<0.01).3.The IHC results showed that for each of the treatment and placebo group,there was significant decrease on the expression of COX2 and Ki67 between the baseline and one year later(P<0.01),and the decrease on the expression of COX2 and Ki67 for treatment group was significantly lower than the placebo group(t values 3.67 and 5.56,P<0.01).For treatment group,the expression of TMEM16A was significantly decreased one year later(P<0.01).The expression of TMEM16A?COX-2?Ki67 proteins all show strong positive on high grade intraepithelial neoplasia and mucosal carcinoma.4.Comparisons of COX2 and Ki67 negative transformation for both of the treatment and placebo groups between baseline and one year after:after one year,the number of negative transformation of COX2 or Ki67 was increased for each group,the number of negative transformation of COX2 or Ki67 in treatment group was more than that in placebo group(P<0.01).5.The IHC results showed that compared to normal colorectal tissues,the expression of TMEM16A,HER2 and EGFR at colorectal adenocarcinoma was significantly higher(P<0.01).6.The positive rate of TMEM16A expression at colorectal adenocarcinoma was 36.94%(58/157),the Chis square or Fisher analysis results showed that tumor grade?TNM group?tumor budding?lymphovascular invasion and nerve invasion were significantly associated with the positive expression of TMEM16A protein(P<0.05).Especially there was more overexpression rate of TMEM16A at mucinous carcinoma than non-mucinous carcinoma(63.64%)(P<0.01).7.The positive rate of HER2 expression at colorectal adenocarcinoma was 32.48%(51/157),the Chis square or Fisher analysis showed that tumor grade,TNM group,the depth of tumor invasion was significantly associated with the positive expression of HER2 protein(P<0.05).8.The chis square or Fisher analysis showed that gender?location?tumor grade?tumor budding and TNM group was significantly associated with the co-expression of TMEM16A/HER2(P<0.05);the co-expression of TMEM16A/EGFR was significantly associated with grade?TNM group?tumor budding,lymphovascular invasion,nerve invasion and mucinous cancer(P<0.05).9.The positive rate of TMEM16A/EGFR co-expression at colorectal adenocarcinoma was 36.31%(57/157),the Spearman's rank correlation coefficient was 0.244(P<0.01)for the correlation between TMEM16A and EGFR expression.10.Bivariate and multivariate logistic regression were performed for the association analysis between the TMEM16Aexpressions,TMEM16A/HER2,TMEM16A/EGFR co-expression and different clinic pathology factors:.At the bivariate analysis,the higher of the TMEM16A proteins expression,the higher of tumor grade,the higher likelihood for tumor budding,lymphovascular invasion,nerve invasion and mucinous cancer.the results for TMEM16A/EGFR co-expression were similar with the results from TMEM16A expression only.The results from multivariate analysis showed that tumor budding and the appearance of mucinousl carcinoma were significantly associated with the overexpression of TMEM16A protein.While the results for bivariate and multivariate simultaneously indicated that female?mucinous adenocarcinoma patients?higher grade tumor budding had a higher likelihood to express TMEM16A/HER2 proteins.11.The results from IHC and Western blotting showed that the positive expression of TMEM16A was higher at tissues from non-small cell lung cancer(NSCLC)than the expression from the normal adjacent tissues.12.The amplification of TMEM16A mRNA was detected by using real-time fluorescence quantitative PCR at NSCLC.13.The overexpression of TMEM16A was related to more advanced NSCLC stage,always accompanied by the overexpression of EGFR.14.The results of NSCLC patients follow up showed:the higher of the TMEM16A expression,the higher of the tumor recurrence rates(39%for TMEM16A expression,and 16%for no TMEM16A expression).(P<0.05)Conclusions:(1)Jianpi-Huayu decoction reduced the recurrence after endoscopic resection of colorectal adenoma.The underlying molecular mechanisms may downregulate the expression of TMEM16A,COX2 and Ki67 at colorectal mucosa epithelial cells,and decrease the proliferation and malignant transformation of tumor cells.(2)TMEM16A expression were independent predictor for tumor budding?lymphovascular invasion etc,the poor prognosis of colorectal adenocarcinoma.The signal transduction pathway of TMEM16A may crossly linked with the signal transduction pathways of HER2 and EGFR.(3)TMEM16A and EGFR may be correlated to the late stage prognosis of NSCLC patients,and could be used to predict the risk of recurrence after surgical removal of the tumors.(4)In summary,TMEM16A expression could induce the development,prognosis and invasion and metastasis at precancerous lesions and malignant epitheliums through various signal transduction pathways on diversified cancers.TMEM16A would be used as a potential prognosis and combined target therapy marker for cancer. |
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