| Colorectal cancer is commonly digestive tract tumor,accounting for the third place in all gastrointestinal tract cancer. Colorectal cancer often locates in the rectum and the junction of rectum and signoid colon,accounting for 65% and occours in the age afte 40 years old.Among the patients, Male to female ratio is 1.27 to 1. The higest incidence of the age groups is among forty years and fifty years old.Most records show that the incidence of colorectal cancer has a increasing trend and the curative effect wasn' t optimistic with increasing of people's living level and changing of dietary structure.The five-survival rate has kept among 50% and 60% in the past 25 years.Moreover,Recurrence and distant metastasis are the main reason of the adverse healing of colorectal cancer.The related basisc research on colorectal cancer is imperative, which provides guidance for clinical diagnosis and treatment. Our previous study suggested that filamin A was closely related with the occurrence and development of breast cancer prostate cancer and malignant cancer.Recently along with research went thorough, it has been acknowledged that filamin A plays an important role in the formation and development of colorectal carcinoma,which may be a new molecular marker of detection of the occurrence, formation, recurrence and metastasis in colorectal carcinoma,furthermore, deep research on the mechanisms of the antineoplastic effect of filamin A provides theoretical basis for Establishment of individual and targeted theraphy of colorectal carcinoma in clinic.In the previous research, haematoxylin-eosin staining and immunohistochemistry(SP) were employed to sixty samples,(people's uterine tissre as negative control and phosphate buffer solution instead of anti-one as positive control),the results of which showed that the expressions of FLNa were lower in the colon cancer groups than those in the normal groups.Also,the research had proved that FLNa had low expression in pathological state and had inhibitory effect on the occurence of colorectal adenocarcinoma, moreover, the expression level of FLNa had negative correlation with invasion of the colorectal cancer and FLNa had significant inhibitory effect on the invasion and metastasis of human colon carcinoma cell,so it would become be a new promising criterion for diagnosis cure and prognosis of colon and rectum tumor. In addition, in the previous research, detection of RT-PCR and Western in colorectal and rectal adenocarcinoma tissue of resection (60 samples ) and normal tissue had proved that positive expression rate of MMP-9 in colorectal adenocarcinoma was higher than that in normal tissue. The expression level of MMP-9 had negative correlation with the expression of FLNa in colorectal adenocarcinoma and had some correlation with TNM stage,liver shift,lymph node metastasis and intestinal infiltration depth. The experiment would study in depth that filamin A' s effect on invasion and metastasis of human colon carcinoma cell line SW480 in vitro and preliminarily investigate the mechanisms of the antineoplastic effect of filamin A to obtain new methods to diagnosis and treatment of colorectal and rectum cancer by cell transfection,RT-PCR,Western blot detection,MTT detection,wound healing assay,transwell chamber experiment,and matrigel invasion assay .Objective: To explore the inhibitory effect of Filamin A(FLNa) on invasion and metastasis of human colon carcinoma cell line SW480 and investigate its mechanisms of the action.Methods:1 In vitro culture of human colon carcinoma cell line SW480 in vitro2 Expressing vector pcDNA3.1/V5-His-TOPO/ FLNa was transfected into SW480 cells by lipofectamine protocols.After G418 selection,the SW480/FLNa cells with expressing of FLNa stably were obtained,all the same time, expressing vector pcDNA3.1/V5-His-TOPO/ without FLNa was transfected into SW480 cells by lipofectamine protocols,so the control group cells were obtained. 3 Expression of FLNa in tumor cells including the group wild cells with not transferred dyes (SW480),the group cells not expressing vector pcDNA3.1/V5-His-TOPO/ FLNa (SW480/pcDNA3.1) and expressing vector pcDNA3.1/V5-His-TOPO/ FLNa (SW480/FLNa) was confirmed by reverse transcription polymerase chain reaction(RT-PCR) and Western blot.4 Cell proliferation in vitro was assessed by MTT assay and cell growth curve was drew to detect cell 's survival and growth.5 Inhibitory effect of filamin A on SW480 cells was observed by wound healing assay,transwell chamber experiment,and Matrigel invasion assay.Results:1 The recombinant vector (pcDNA3.1/V5-His -TOPO/FLNa)was correctly transfected into SW480 cells by liposome in human colon carcinoma cell line SW480.2 The expressions of FLNa mRNA were higher in the SW480/FLNa cells than those in the wild SW480 cells, (1.27±0.03) vs (0.14±0.02) , P < 0.01 respectively.3 The expressions of FLNa protein were higher in the SW480/FLNa cells than those in the wild SW480 cells, (1.18±0.03) vs (0.25±0.02),P<0.05,respectively.4 Growth Curve of three groups were almost overlap and proliferation ability of three groups had no difference.5 Results of wound healing assay: the migration velocity in group SW480 was significantly faster than that in group SW480/FLNa,but the difference was insignificant between group SW480 and group SW480/pcDNA3.1.6 Results of transwell chamber experiment and matrigel invasion assay: The number of transwell cells in group SW480/FLNa was decreased significantly than those in group SW480 (P <0.05).,but no difference was found between in group SW480/pcDNA3.1 and in group SW480/FLNa.Conclusion:1 FLNa has significant inhibitory effect on the invasion and metastasis of human colon carcinoma cell line SW480. 2 FLNa has a high correlation with the occurrence,development and invasion of colorectal carcinoma, and high expression of FLNa protein has significant inhibitory effect on the invasion and distant metastasis of colorectal carcinoma.
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