Correlation Of Lymphatic Vessel Density With HMGB1and TAMs In Cervical Carcinoma

Objective①To confirm the expression of HMGB1,CD68(TAMs Specific markers) and D2-40(LVD Specific markers) in Cervical squamous cell carcinoma, and to analyze the expression level of three indexes by Statistical methods.②To investigate the relationship between HMGB1, TAMs and LVD in patients with clin-ical pathological characteristics.③To investigate the correlation between HMGB1, T AMs and LVD in Cervical squamous cell carcinoma and its effect on the prognosis of the patients, which provide important experimental evidence for clinical prognosis. Methods①to collect accept radical operation of cervical carcinoma, confirmed by pathology in patients with idiopathic CSCC paraffin embedded tumor tissue specimens from93cases in Tianjin Medical University Cancer Institute and Hospital from2007in January to2008in December. All patients without preoperative chemotherapy, radiotherapy or other therapy history. Patients aged from25to68years, the median age was44years old.The age over44years old was51cases,42cases were under44years of age; pathological classification for Cervical squamous cell carcinoma:high-middle differentiation, low differentiation were65cases,28cases;20cases were with lymph node metastasis,73cases were of metastasis without occurred.14cases were tumor diameter greater than4cm,79cases were less than or equal to4cm. The depth of invasion:shallow muscle layer (less than or equal to1/2) in45cases, deep muscle layer (greater thanl/2) in48cases; FIGO stage:5cases of LA stage, IB stage33cases, IIA26cases, IIB in29cases.In addition,to collect the normal cervical squamous epithelium tissue samples was as control study from40cases (for the myoma of uterus hysterectomy).②Immunohistochemical method was used to detect the expression of HMGB1, CD68and D2-40in93cases of CSCC and40cases of normal cervical squamous epithelial tissues, and HMGB1counts, TAMs counts and counts of lymphatic vessel were assessed after the outcomes.By SPSS17.0statistical software, t test was used for continuous variables,χ2test was used for categorical variables; Kaplan-Meier method and Cox regression were used for survival analysis; correlation analysis was carried out by using Spearman rank correlation analysis. Results①Three index for HMGB1, CD68and D2-40were higher expression in squamous cell carcinoma of the uterine cervix, and its expression level was significantly higher than that of normal cervical squamous epithelial cells, it was statistically significant for the difference (P<0.05).②Expression of SP immunohistochemical method was used to detect HMGB1, CD68and D2-40in93cases of CSCC and40cases of normal cervical squamous epithelial tissues, and HMGB1count, TAMs count and counts of lymph vessels.HMGB1expression and age, tumor size, depth of invasion, lymph node metastasis had a relationship with FIGO stage (P<0.05), but not relationship with the degree of differentiation (P>0.05). TAMs count and diameter of tumor, histologic differentiation, invasion depth, lymph node metastasis had a relationship with FIGO stage (P<0.05), and without relationship to age (P>0.05). LVD and tissue differentiation, invasion depth, lymph node metastasis had relationship with FIGO stage (P<0.05), and not with age, diameter of the tumors (P>0.05).③HMGB1and expression of TAMs increased, LVD increased; HMGB1and expression of TAMs decreased, LVD decreased. After the Spearman rank correlation analysis, positive correlation between HMGB1expression, TAMs count and LVD.④High expression of HMGB1, LVD high expression group survival time (54.45±11.275months) was significantly lower than that of LVD low expression group survival time (61±5.904months), the difference was statistically significant between the two groups (t=2.138, P=0.037).High expression of TAMs, LVD high expression group survival time (53.44±13.721months) was significantly lower than that of LVD low expression group survival time (61.57±7.659months), the difference was statistically significant between the two groups (t=2.432, P=0.019).Single factor analysis showed that, the group of lymph node metastasis in3years,5year survival rates were75%,36.4%, significantly lower than the group without lymph node metastasis (98.6%、87.8%)(x2=32.925, P<0.01) High expression of HMGB1groups were3years,5years survival rate (89.5%,61.4%) was significantly lower in patients with low expression group (100%,96.8%), the difference was statistically significant between the two groups (χ2-11.366, P<0.01).The mean counts (mean47.656) of TAMs as cut points are divided into two groups, the high expression group of3years,5years survival rate (89.6%,60.3%) was significantly lower in patients with low expression group (97.8%,94%), the difference was statistically significant between the two groups (χ2=11.448, P<0.01).The mean LVD (mean10.623) cut points are divided into two groups, the high expression group were3years,5years survival rate (92.3%,61.3%) was significantly lower in patients with low expression group (95.1%,95.1%), the difference was statistically significant between the two groups (χ2=11.251, P<0.01).Multivariate analysis showed that the expression of lymphatic metastasis and HMGB1as independent prognostic factors (P=0.001, P=0.043), while TAMs and LVD are not independent prognostic factors.Conclusion①HMGB1proteins and TAMs were highly expressed in CSCC.Patients who exhibit increased HMGB1expression or increased TAMs count consequently show enhanced LVD expressions,increased lymph node metastasis,and poor prognosis.②HMGB1and lymph node metastasis were independent prognostic factors in patients with Cervical squamous cell carcinoma, which provides an important evidence for clinical prognosis. It is of great significance for the follow-up in-depth study of Cervical squamous cell carcinoma.