| OC(ovarian cancer) is the one of the most common gynecologic malignant tumors and its mortality rate ranks top of all the gynecologic cancers. EOC(epithelial ovarian cancer) occupies 50%-70% in OC and is the most important pathological type, among which ovarian serous carcinoma appears most. Lack of specific symptoms in early time, late recurrence and high metastasis cause the high mortality rate of OC. Therefore it is importantly significant to improve the early diagnosis rate and prevent the recurrence and metastasis for improvement of patients’ lifetime and prognosis. Now the clinical methods for the diagnosis of early OC are not clear. Researchers have shown that the combined detection of CA125, HE4 and other tumor markers can improve the early diagnosis rate of OC. Besides, HK6, HK10 and other emerging tumor markers are also helpful for the early diagnosis of OC. However there is not a specific method for early OC. The clinical treatment of OC usually contains operation and chemotherapy. In recent years new technology including molecular therapy, gene therapy and immunotherapy is achieving increasing success, especially immunotherapy is becoming more and more popular. Therefore to find an effective target for diagnosis and treatment of is the main problem for ovarian cancer.The transfer form of ovarian cancer mainly includes direct spread, celiac planting and lymphatic metastasis. Tumor cells break away from primary focus into the abdominal cavity, adhere to the abdominal viscera and invade capsule, causing the mass propagation of tumor cells and formation of new cancerous focus. This process is regulated by oncogene and anti-oncogene, EMT(the epithelium-interstitial transformation), anti- anoikis and immune escape etc.This behavior which unlimitedly proliferate and multi-directional differentiate of tumor cells and embryonic stem cells are similar, so people speculated that the tumor is a kind of "atavism". Thus it can be seen that between the embryonic development and tumor occurrence and metastasis there might be common regulatory mechanism. Some embryonic factors might have played an important role in tumor occurance and metastasis.FOXC2(forkhead box C2,) is an embryonic transcription factor belonging to the FOX family, which was first found in LD(Lymphoedema- Distichiasis Syndrome). Studies have shown FOXC2 played an important role in lymphatic vessel formation and the normal development in embryonic period. Mouse embryos with FOXC2 knock-out were deficient in normal form and function of lymphatic vessels. And several researches have proved FOXC2 takes effect in invasion and metastasis of various tumors. Studies have shown that FOXC2 can promote the invasion and metastasis of colorectal cancer and it can also promote human breast cancer cell in invasion through EMT. There are studies about that FOXC2 can promote the metastasis of cervical cancer trough tumor angiogenesis. Another study shows that FOXC2 can help cervical cancer cells and endometrial cancer cells migrate and metastase, and its mechanism may be related with lymph angiogenesis. Nevertheless the relationship between FOXC2 and OC is still uncertain. This experiment is mainly about EOC, and tries to find out how FOXC2 acts in EOC through immunohistochemical and RNAi technology, laying a good foundation for building the mouse ovarian cancer homograft model and providing a new target for the early diagnosis and treatment of OC. Objective:Observe the expression mode of FOXC2 in human OC and how biological behavior changes of mouse gland cancer cells ID8 after silencing FOXC2 gene, to investigate how FOXC2 influent the proliferation and invasion of OC and look for a new target for early diagnosis and treatment. Methods:1)Observe the distribution of FOXC2 in ovarian epithelial cancer, borderline ovarian tumor and normal ovarian tissues with immunohistochemical method and the clinical samples are all from the first hospital of Jilin University.2)Design the target gene sequence of mus Foxc2-sh RNA sequence, construct the recombinant plasmid and package lentivirus through cotransfecting with 293 T cells and last test the titre.3)Infect ID8 cells with lentvirus vector to detect the inhibition of Foxc2 expression with real time PCR and Western Blot4)Observe the changes of cell proliferation ability of ID8 after infection with lentivirus through CCK8 method.5) Observe the changes of cell invasion ability of ID8 after infection with lentivirus through transwell test. Results:1) FOXC2 is positive in ovarian epithelial cancer and mainly expressed in nucleus; FOXC2 is positive in the cell wall of endocrine gland in serous borderline tumor;FOXC2 gene is also expressed in blood vessels of tumor stroma in ovarian cancer as well as borderline tumor2) The result of recombinant plasmid DNA sequencing is the same with expected sequence proving the plasmid is successfully reconstructed. The titre shows the lentivirus vector is successfully built.3) Silencing Foxc2 can inhibit the expression of Foxc2 m RNA and the inhibition rate is respectively 71.36% and 56.3%(P<0.05); Silencing Foxc2 can also inhibit the expression of Foxc2 protein, in which the first experimental group was inhibited most obviously.4) After 48 h of silencing Foxc2 the proliferation rate of ID8 cell begins to slow down, after 72 h and 96 h the cell proliferation rate slowed down more obviously(P<0.05).5) After silencing Foxc2 the number of invasive cells in experimental group was declined 72.2% compared to control group(P<0.05). Conclusions:1) FOXC2 expression is positive in both ovarian epithelial cancer and borderline tumor, and expressed in both borderline tumor and cancer, proving that FOXC2 is involved in the occurrence and metastasis of ovarian cancer.2) The test constructs successfully the FOXC2 silencing lentivirus vector.3) Silencing Foxc2 can inhibit the proliferation and invasion of ID8 cells.4) FOXC2 can be a new target for the clinical diagnosis and treatment of OC. |
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