| Background: The prognosis of esophageal carcinoma, one of the most commonmalignancies, is still poor today. Even the treatment of esophageal carcinoma withtraditional Chinese medicine (TCM) has a very long history, its molecular biologicalmechanism has not been understood clearly. In this paper, we studied the moleculareffectiveness of Aidi injection, a traditional Chinese Compound Medicine injection, onpatients with esophageal squamous cell carcinoma (ESCC). We gave Aidi injection topatients with ESCC prior to surgery, and then observed various over 24 indexes ofimmunohistochemistry (IHC) and apoptosis by tissue microarray of the cancer tissueafter surgery in comparison with the control group, in order to illuminate the molecularbiological mechanism of Aidi injection on patients with ESCC in pre-operation andprovide the possible theory for combined therapy of ESCC.Methods: 46 ESCC patients were grouped into experiment and control panel, each ofwhich included 23 subjects. Patients of the experiment group were given 80ml/d,intravenous injection of Aidi for 7 days before surgery, while patients of the controlgroup were not given any Aidi injection. Other related therapeutic medicines were assame in both groups. After completing ordinary H&E staining detection, tissuemicroarray (TMA) was prepared and performed using immunohistochemistry (IHC) andterminal deoxynucleotide transferase-mediated dUTP nick end labeling (TUNEL)method. 24 antibodies were observed by immunohistochemistry staining. Theseindexes were divided into four groups:①multidrug resistance associated indexes,including LRP, MRP, MGMT, TS, P-gp, TopoⅡa and GST-p;②indexes related totumor proliferation, cell differentiation, metastasis and prognosis, including Ki-67, ER,PgR, COX-2, TIMP-1, TP and HSP70;③indexes related to oncogene expression,including c-fos, C-myc, p53, c-erbB-2, nm23 and MDM2;④indexes related toapoptosis, including Bcl-2, Bax, NFκBP50 and Fas. While ESCC apoptosis wereobserved by TUNEL method. In addition, directly sequencing methods was used to analyse the sequence mutation of p53 exon 5-8.8 biopsy tissues of patients in the Aidiinjection group and 3 biopsy tissues in the control group were examined.Results: There was no significant statistical difference between the two groups in sex,age, tumor sites, tumor size, differentiated degree, infiltration degree or lymph nodemetastasis.①Regarding to multidrug resistance associated indexes, the positiveexpression rates of LRP、MRP、MGMT、TS、P-gp、TopoⅡa、GST-p in the Aidiinjection group were 26.1％、34.8％、69.6％、69.6％、34.8％、43.5％and 87.0％,respectively, while those in the control group were 39.1％、47.8％、52.1％、82.6％、39.1％、47.8％and 87.0％respectively. There was no significant statistical differencediscovered in these seven indexes between the two groups (P＞0.05).②Regarding toindexes related to tumor proliferation, cell differentiation, metastasis and prognosis, thepositive expression rates of Ki-67, ER, PgR, COX-2, TIMP-1, T P, HSP70 in theexperimental group were 65.2％, 13.0％, 13.0％, 87.0％, 34.8％, 69.6％and 8.7％,respectively, while those in the control group were 43.5％, 17.4％, 4.3％, 78.3％,65.2％, 60.9％and 13.0％respectively. There was no significant statistical difference inthese indexes between two groups(P＞0.05).③Regarding to indexes related to oncogeneexpression, the positive expression rates of c-los, C-myc, p53, c-erbB-2, nm23 andMDM2 in the experiment group were 69.6％, 0％, 39.1％, 56.5％, 69.6％and 52.2％,respectively, while in the control group were 52.2％, 4.3％, 56.5％, 56.5％, 87.0％and47.8％, respectively. There was no significant difference discovered in statistics betweenthem (P＞0.05).④Regarding to indexes related to apoptosis, the positive expression forBcl-2、Bax、NFκBP50 and Fas in the experiment group were 39.1％, 56.5％, 47.8％and 47.8％respectively, while in the control group were 78.3％, 17.4％, 87.0％and39.1％respectively. The positive rates of Bcl-2 and NFκBP50 in the experiment groupwere significantly lower than those in the control group (P＜0.05). The rate of Bax inthe experiment group was significantly higher than that in the control group (P＜0.05).There was no significant difference discovered in the expression of Fas between twogroups (P＞0.05). The ratio of Bax/Bcl-2 in the experiment group(1.22±0.72) wassignificantly higher than that in the control group(0.47±0.65) (P＜0.01). There was apositive correlation between NFκBP50 and Bcl-2 (r_s=0.358,P＜0.05). In addition, thepositive rate of apoptosis in the experimental group(69.6％) was significantly higher thanthat in the control group(8.7％) (P＜0.01) by TUNEL method. There was negativecorrelation between NFκBP50 and positive rate of apoptosis (r_s=-0.297, P＜0.05). ⑤Correlation analysis among these indexes: there was no statistical correlation betweenNFκBP50 and MRP (r_s=-0.076, P＞0.05)、P-gp (r_s=-0.044, P＞0.05)、Ki-67(r_s=-0.044, P＞0.05)、COX-2 (r_s=-0.048, P＞0.05)or p53 (r_s=0.016, P＞0.05); whilethere were positive correlation between ER and PR (r_s=0.299, P＜0.05), MRP andLRP (r_s=0.641, P＜0.01)、p53 and Ki-67 (r_s=0.528, P＜0.01)、p53 and MDM2(r_s=0.348, P＜0.05). (6) The ratio of mutation on p53 exon5-8 in the Aidi injectiongroup was 50％(4/8), while those in the control group was 33.3％(1/3). The total ratio ofmutation and polymorphism on p53 exon7-8 in the Aidi injection group was 100％(8/8),while in the control group was 66.7％(2/3).Conclusions: Aidi injection may interfere with ESCC prior to surgical operation throughthe following pathways:①Aidi injection could downregulate the expression of Bcl-2and increase the ratio of Bax/ Bcl-2 to improve the apoptosis of ESCC.②It hasshowed some statistical correlation with the blockage of NFκB pathway. Aidi injectioncould induce apoptosis of ESCC by inhibiting the activity of NFκB thendownregulating the expression of Bcl-2.③The incidence of esophageal cancer isassociated with a variety of molecular mechanisms that affect each other. So performingAidi injection to cancer patients as a pre-surgical clinical treatment has molecularpathologic mechanisms-effects. It is feasible to use tissue microarray technique, which isa rapid, economic and accurate method to screen clinical tissue specimens on a largescale. Also, Aidi injection could not decrease the detection rates of mutation andpolymorphism on p53 exon5-8. |
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