| Objective:The exosomes in laryngocarcinoma and hypopharyngeal carcinoma are extracted and discussed in terms of their clinical significance.Materials and methods:A total of 22 cases of preoperative peripheral blood have been collected from the patients with head and neck cancer,including 11 cases of laryngeal cancer patients and 11 cases of hypopharyngeal cancer patients.Meanwhile 11 additional cases of normal people have also been collected for comparisons.All follow-up studies based on these cases have proved by Academic Institutional Review Board(AIRB)with an informed consent form signed by patients and their families.1.The exosomes were extracted from the collected bloods of laryngocarcinoma.hypopharyngeal carcinomapatients and normal persons by System Biosciences(EXOTC10A-1).2.The shape of exosomes were observed through the transmission electron microscope.3.Nanoparticle Tracking Analysis(NTA)has been used to make a count and a particle diameter measuring for exosomes.4.Bicinchoninic acid(BCA)has been used to identify the protein concentration of exosomes.5.The expression of protein were detected by making advantage of the Western blot.6.The expressiones of exosomes in patients having head and neck cancer(laryngocarcinoma?hypopharyngeal carcinoma)22 cases and normal persons 11 cases are detected.There are 11 cases of laryngocarcinoma patients and 11 cases of hypopharyngeal carcinoma,all of which are validated by the biopsy under laryngoscopy.7.The statistical software,SPSS 19.0,had been adopted.Results:1.It is possible that separate out the exosomes from the supernatant liquids of laryngocarcinoma and hypopharyngeal carcinoma patients.The separated exosomes under transmission electron microscope appear to be characterized by a cup-shaped structure,having 30-100 nm diameters.2.Based on the observation by transmission electron microscope,the volumes of exosomes from the blood serums of laryngocarcinoma patients as well as the hypopharyngeal carcinoma patients are larger than the ones from normal people.3.The protein concentration of exosomes from the laryngocarcinoma and hypopharyngeal cancer patients are significantly greater than the ones from normal people(p<0.05).From the ROC curve in terms of the protein concentrationbetween the normal cases and cancer cases(including laryngeal cancer and hypopharyngeal cancer),the area under curve(AUC)is 0.926±0.046,and the Youden index reaches the maximumvalue of 0.864when the protein concentration of exosomes is 2.8842.Based on this,it can be found that the diagnosticcritical value of protein concentration of exosomes is 2.8842,the diagnostic sensitivity is 86.4%,and the specificity is 100%.Hence,the protein concentration of exosomes from the peripheral blood can be considered as one of the markers for early diagnosis of laryngocarcinoma and hypopharyngeal carcinoma.4.The CD63and CD81proteins had been detected in the exosomes from the blood serums of normal people,laryngocarcinoma and hypopharyngeal carcinoma patients,while the Calnexin protein were failed to be detected.5.The particle concentrations of exosomesin the blood serums are not significantly differentin the normal people,laryngocarcinoma and hypopharyngeal carcinoma patients(p>0.05).Compared with the case of normal people,the particle diameterof exosomes in the blood serumsfrom laryngocarcinoma patient appears larger,yet without an evidentdifference between them.Meanwhile,the particle diameter of hypopharyngeal carcinoma patient is significantly larger than that of normal people(p<0.01).Based on the ROC curvein terms of the particle diameter between the normal cases and laryngocarcinoma cases,it is found that the AUC is 0.661 ±0.118,and the maximal Youden index is 0.181.Additionally,the diagnostic sensitivity and the specificity are not high,which are 63.6%and 54.5%,respectively.While by the ROC curve in terms of the particle diameter between the normal cases and hypopharyngeal carcinoma cases,it is found that:the AUC is 0.979±0.024,the Youden index reaches the maximal value of 0.808 when the particle diameter is at its critical value of 162nm,the diagnostic sensitivity is 100%,and the specificity is 81.8%.Based on this,theparticle diameter of exosomes from the peripheral blood can also be considered as one of the markers for early diagnosis of hypopharyngeal carcinoma.Conclusion:The exosomes from the blood serums of laryngocarcinoma and hypopharyngeal carcinoma patients have a great potential to be as the tumor marker,which will be of an immense help to the early diagnosis,surgical effect,prognosis and postoperative follow-up. |
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