Study Of The Relationship Between Neoplasm Of Lymphatic Vessel And Metastasis Of Lymph Node In Early Stages Of Cervical Cancer

Objection: Cervical cancer is the most common gynecological malignant tumor. Lymphatic metastasis is the most important cause for treatment failure and death of cervical cancer. This research studied the relationship between neoplasm of lymphatic vessel and metastasis of lymph node in early stage of cervical cancer, in order to find the critic index for prediction of lymphatic metastasis, and to provide the new therapy target for suppression lymphatic metastasis and improvement the treatment effect.Methods:(1) Using Transmission electronic microscopy and Optical microscopy to observe the distribution and morphologic character of micro lymphatic vessels in the region of intra-cancer, para-cancer, normal cancer and sentinel lymph nodes (SLN) of cervical cancer. Analysis the relationship between metastasis of sentinel lymph nodes and neoplasm of lymphatic vessels, lymphatic vessels total area (LVTA), lymphatic vessels invasion (LVI) and opening connection of lymphatic endothelia cells.(2) Using Immunohistochemistry (IHC) and Real time Reverse transcription polymerase chain reaction (RT-PCR) to detect the expression of VEGF-C/D in the tissue of intra-cancer, para-cancer, normal, SLN and non-SLN. And analysis the relationship of the expression of VEGF-C/D and neoplasm of lymphatic vessels, invasion of lymphatic vessels and metastasis of SLN, exploration the mechanism of neoplasm of lymphatic vessels, invasion of lymphatic vessels and metastasis of SLN. (3) Analyzing the detection of LVI-IHC and SLN biopsy, and the value of combining LVI-IHC and SLN for predicting the metastasis of lymph nodes in cervical cancer.Results:(1) Distribution and morphologic characters of lymphatic vessels of cervical cancer: There were few collapsed lymphatic vessels in the region of cervical cancer tissue and could hardly be counted. The density of lymphatic vessels (LVD) in para-cancer region was significantly higher than that in normal region (p=0.000). LVTA in para-cancer region was significantly lower than that in normal region (p=0.002). The LVD and LVTA of metastasis lymph nodes were significantly higher than that of lymph nodes without metastasis. There were cancer cells invasion into lymphatic vessels in para-cancer region, but no cancer cells invasion into lymphatic vessels in normal region.(2) Relationship between invasion of lymphatic vessel in para-cancer region and metastasis of SLN: There were 35.92% (18/51) para-cancer regions of cervical cancer having lymphatic vessel invasion, among which there were 16 cases having positive SLN. HE detected 13 cases of positive SLN, its correct rate for predicting SLN was 94%(47/50), false negative was 18.75%(3/16). While IHC detected 14 cases of positive SLN, its correct rate for predicting SLN was 96%(48/50), false negative was 12.50%(2/16). There were uniformity between invasion of lymphatic vessel in para-cancer region and metastasis of SLN (McNemar Test p=0.500). Using invasion of lymphatic vessel in para-cancer region to predict metastasis of SLN, the positive predictive value was 83.33%(15/18),negative predictive value was 93.93%(31/33),correct rate was 90.19%(46/51).(3) The expression of VEGF-C in cervical cancer tissue and its clinical significance: The positive rate of VEGF-C in cervical cancer was 70.59%(36/51),in para-cancer region was 68.63%(35/51), in 17 cases of metastasis lymph node(among them 16 was SLN) was 94.12%(16/17). There was no VEGF-C expression in normal cervical tissue and lymph node without metastasis. The expression of VEGF-C in different tissues has significant difference( p=0.000). The metastasis rate was 44.44%(16/36) in VEGF-C positive group , which was significant higher than VEGF-C negative group 13.33%(2/15). The invasion of lymphatic vessel was 44.44%(16/36)in VEGF-C positive group, which was significant higher than VEGF-C negative group 6.67%(1/15). The positive expression of VEGF-C 47.22%(17/36)was significantly higher than negative expression of VEGF-C 13.33%(2/15)in Stage II of cervical cancer(p<0.05). Both LVD and LTA of VEGF-C positive group were significantly higher VEGF-C negative group(p<0.05).(4) The expression of VEGF-D in cervical cancer tissue and its clinical significance: The positive rate of VEGF-D in cervical cancer was 60.78%(31/51),in para-cancer region was 56.86%(29/51), in 17 cases of metastasis lymph node(among them 16 was SLN) was82.35%(14/17). There was no VEGF-D expression in normal cervical tissue and lymph node without metastasis. The expression of VEGF-D in different tissues has significant difference( p=0.000). The metastasis rate was 51.61%(16/31) in VEGF-D positive group , which was significant higher than VEGF-D negative group 3.33%(1/30). The invasion of lymphatic vessel was 48.39%(15/31)in VEGF-D positive group, which was significant higher than VEGF-D negative group 10%(3/30)(p<0.05).(5) The expression of VEGF-C/DmRNA: Using Real time quantity of RT-PCR to detect the expression of VEGF-C mRNA. The expression of VEGF-C mRNA was significantly higher in intra-cancer, para-cancer, and metastasis SLN than that in normal cervical cancer (P<0.01)and non-SLN without metastasis(P<0.01). The expression of VEGF-C mRNA was significantly higher in metastasis SLN than that in intra-cancer and para-cancer region(P<0.05). The expression of VEGF-D mRNA was significantly higher in intra-cancer, para-cancer and, metastasis SLN than that in normal region(P<0.01)and non-SLN without metastasis(P<0.05). The expression of VEGF-D mRNA was significantly higher in metastasis SLN than that in intra-cancer, but had no significantly different with para-cancer region(P>0.05). There were significantly difference in expression of VEGF- C/D mRNA in LN metastasis, LVI and depth of cancer invasion(P<0.05).(6) The success rate of combining 99mTc and dye to detect SLN was 98.04%(50/51). There were 141 SLN that were detected. HE detected 35(24.82%) positive LN, IHC detected 42(29.79%)positive LN. There were no significant difference between them(p=0.283).(7) There were 18 cases having LVI, among them there were 15 cases of metastasis of SLN. Using LVI-IHC to predict metastasis of SLN, the sensitive rate was 93.75% (15/16), specific rate was 90.32% (28/31), correct rate was 98% (49/50), false negative rate was 6.25%(1/16). Combining LVI-IHC and SLN detection to predict metastasis of lymph node, the sensitive rate was 100%(16/16), specific rate was 90.32% (28/31), correct rate was 100%(50/50), false negative was 0%(0/16).(8) The influence of clinical stages on SLN: There was only 1 case which was failed to detect SLN. The diagnosis correct rate was 100%(25/25)in stage Ib1, 100%(7/7) in stage Ib2, 88.89%(16/18)in stage IIa, false negative rate was 16.67%(2/12).Conclusion:(1) The neoplasm of lymph vessels of cervical cancer happened in para-cancer region. The lumen of neo-lymphatic vessel was small. There maybe existed two main methods for lymph node metastasis: tumor cell pass through the open connection of lymphatic vessel wall, or tumor cell distroy lymphatic vessel of para-cancer region. LVI-IHC of para-cancer region may become a sensitive index of predicting metastasis of lymph nodes.(2) Both VEGF-C and VEGF-D were up regulated in cervical cancer tissue, and they have synergistic effect. Both VEGF-C and VEGF-D could induce neoplasm of lymphatic vessel in para-cancer region, and improve invasion of lymphatic vessel, and increase metastasis of lymph node. Maybe they take part in the process of happen and development of cervical cancer, and act important effect in lymph node metastasis. They may become a significant index for lymph node metastasis and treatment target for suppression lymph node metastasis and improvement treatment effect.(3) The combining detection of LVI-IHC and SLN for Ib cervical cancer, may improve the sensitivity of diagnosis for lymph node metastasis and decrease the false negative.