Expressions And Significance Of DNA Ploidy, Ki67 And CD44v6 In Conventional Pathologic Non-metastasis Lymph Nodes In The Patients With Early Cervical Squamous Cell Carcinoma

Object:Metastatic spread of cervical carcinoma mainly via the lymphatics.DNA ploidy is the response of the ability of cell proliferation,aneuploid tumors is prone to occur pelvic lymph node metastasis,and the ploid types in metastasized lymph nodes is the same as the original tumor.Ki67 can reflect the activity of cell proliferation accurately as a marker of cervical Carcinoma.CD44v6 participates in the invasion and metastasis of the tumor,the tumor cells of CD44v6 overexpression may acquire "camouflage" of Lymphocytes and then escape from the recognition and killing of human immune system and easily invade the lymphatic system.The purpose of the study was to use DNA ploidy,ki67 and CD44v6 to detect the expression in lymph nodes of early cervical squamous cell carcinoma patients who had received radical hysterectomy and pelvic lymphadenectomy.These lymph nodes had not been invaded by the tumor confirmed by routine histopathological examination.On the other hand,the study was to discuss it's significances in the pelvic lymph nodes micrometastatic Focus.Method:Thirty-two patients withⅠb～Ⅱa period(FIGO stage)cervical squamous cell carcinoma who underwent radical hysterectomy and pelvic lymphadenectomy were enrolled in the present study from Department of Gynaecology,the First Affiliated Hospital of Shanxi Medical University and Shanxi Tumour Hospital.Lymph node and distant metastasis couldn't be found by clinical signs and image examination,without receiving pre-operational radio-chemotherapy, without the history of other cancer and retroperitoneal tumor surgical operation.First of all,3 cases postoperative pathological diagnosis by routine examination of lymph node metastasis were found,9 cases negative metastasis were found from 12 cases without identification of sentinel lymph node(SLN),collected all 156 negative metastasis pelvic lymph node,which were fixed by formalin and embedded by paraffin after surgical excision.Besides,3 cases postoperative pathological diagnosis by routine examination of metastasis were found,17 cases negative metastasis were found from 20 cases with identification of SLN successfully,44 sections fixed by formalin and embedded by paraffin after surgical excision were collected in all negative metastasis pelvic SLN.We apply to flow cytometry(FCM)and immunohistochemistry (IHC)methods,the former was appllied to the determination of the DNA ploidy,and the later was applicable to the determination of expression level of ki67 and CD44v6.The lymph nodes were detected that accrod with selected standards.Each lymph node was cut at three levels with consecutive sections,the 4μm section was for haematoxylin and eosin(HE)staining review, when the review was negative,the rest two were used for IHC(ki67 and CD44v6);then cut the thickness of 60μm sections,which were collected for FCM analysis of DNA ploidy.Results:First,three of 12 patients with cervical carcinoma showed evidence of the possible missed diagnosis metastatic foci by routine pathological examination in the LNs through one or more of the two methods.A total of 156 sections were examined,19 LNs of 3 cases DNA aneuploidy,the rate of positive expression was 9.62%(15/156);15 LNs of 3 cases ki67 is positive,the rate of positive expression was 9.62%(15/156);9 LNs of 2 cases CD44v6 is positive,and the positive expression rate was 8.33%(13/156).one cases was LN positive metastasis patients,in it's respective negative LNs,the above three indicators was different degrees detection of positive expression.One LN in negative metastatic patients was ki67 positive,while other two indexes were negative.Second,in 44 SLNs from 20 patients with early squamous cervical carcinoma,the result of detection by IHC was that there are 7 LNs positive expression in 3 cases.9 LNs of 4 cases DNA aneuploidy,the rate of positive expression was 20.45%(9/44);6 SLNs of 3 cases ki67 is positive,the rate of positive expression was 13.64% (6/44);5 SLNs of 2 cases CD44v6 is positive,the positive expression rate was 11.36%(5/44).Conclusion:Firstly,Compared with traditional histological slice,positive expression can be detected by indexes of DNA ploidy,ki67 and CD44v6,which implied that traditional H&E staining examination may have the possible of misdiagnosis.Secondly,among early squamous cervical carcinoma patients,detection of DNA ploidy by FCM,ki67 and CD44v6 by IHC,which is of certain clinical value of a prompt in the determination of the micrometastases in lymph nodes.The positive rate of DNA ploidy detection is higher than that of ki67 and CD44v6 detection.Finally,it's much easier to find the earliest metastases in SLN than in all LN.