JAK Inhibitor AG490 And Survivin Gene Expression, The Significance In The Ovary Epidermis Cancer Development

Objective: 1 Discusses JAK--STAT( janus kinase --signal transducer and activator of transcription) signal transduction circuit inhibitor AG490 in human ovarian cancer epidermis cell line SKOV3 to survivin (survival element) the gene expression influence and survivin multiplies to the tumour cell, perishes the influence possibility to affect the mechanism weakly.2 Discusses the function of survivin in the ovary epidermis cancer development.3 Discusses AG490 in the anti-tumor ovary aspect research value and the clinical practice prospect.The research indicated that JAK--STAT way widespread participation processes and so on cell's multiplication, differentiation as well as immunity adjustment, are many kinds of cell factors and the growth factor signal transduction important way. In the normal cell, activates STATS is an instantaneous process, but in cancer cell, the STATS protein is activated exceptionally continually, STAT1, STAT2 and STAT6, the STAT1 function mainly suppresses with the growth related, but STAT3 and STAT5 may through suppress apoptosis rate or the induction cell proliferation function participation tumor occurrence development. This experiment medicine AG490 is the JAK enzyme specificity inhibitor, through suppresses the JAK enzyme activeness, but effective blocks its downstream the signal transduction and the duplication activates STAT the activation, affects JAK--STAT signal passage's conveying. But the suppressor cell proliferation receives suppresses, cell's apoptosis rate drop.Methods: Makes the cell culture on behalf of ovary epidermis cancer's cell line SKOV3, Flow cytometry technique examines the situation of the SKOV3 cell'apoptosis rate, finally examines STAT3, p-STAT3 and the survivin expression by the immunohistochemistry. The SKOV3 cell convention raise in contains in 10% embryo cow blood serum's RPMI-1640 culture medium. Adds the logarithm vegetal period's SKOV3 cell with the different density AG490 medicine raises(AG490 medicine density respectively is 0μmol/ L is blank control group, 20μmol/ L, 40μmol/ L, 80μmol/ L, 160μmol/ L), Each group (12 hour, 24 hour, 48 hour) grows in different time with the method of MTT examination cell the situation which inhibit; computation cell growth suppression rate and Situation which cell cycle change and situation cell apoptosis rate change with the flow cytometry; Examines in separately 24 hour control group and the 40μmol/ L AG490 treatment group with the immunohistochemistry group reduction STAT3, p-STAT3 and the survivin expression change situation.Results: Under the microscope observes the control group and various experimental groups cell quantity, the size and the shape change every day. In normal condition: Under the microscope sees the SKOV3 cell to assume the monolayer to grow, the cell long spindle-shaped either the polygon, the nucleus circular or the ellipse, the cytoblastema is translucent. Processes the latter 12 hours after AG490, the cell body starts changes the circle, the cytoblastema passes brightness to drop, the pellet feeling strengthens, the partial cells fall off assume the aerosol shape, and the obvious cell fragment, 24 hour cytomorphology change is more obvious, the most cells assume the circular, falls off combines to make or become the suspended state the cell to increase, 48 hour 80% above cell assumes the circular nearly, in the nutrient fluid has the massive aerosol cells.MTT examines the different density (0-80 umo1/L) AG490 the treatment group SKOV3 cell to select 12 hour, 24 hour, 48 hour growth situation in the different time, presents the phenomenon which the certain extent the suppressor cell multiplication and the induction cell perish weakly, and this kind of inhibitory action assumes the time and the density dependence, After AG490 processing cell,presents the cell multiplication activeness to reduce, the 20, 40, 80, 160μmol/ L treatment group 48 hour cell's growth suppression rate distinction 38.24%, 53.08%, 75.54%, 84.25%, the cell growth reduces speed, the cell number reduces.Flow cytometry technique analysis discover that AG490 causes the SKOV3 mitotic cycle to hinder in the G2/M time, the S time, the G0/1 time cell number to reduce, the cell apoptosis rate increases; Adds may see with the AG490 latter 24 hours obvious cell apoptosis the peak. 48 hours blank control group's apoptosis rate approximately is 5%, 40μmol/ L medicine function group cell apoptosis rate may reach 33%, has the remarkable difference (P<0.05).The immunohistochemistry may see AG490 to be possible through JAK enzyme activeness, thus has suppressed the STAT3 activation, the STAT expression rate from 76.29% rises to 80.66%, non-significance difference (P>0.05), but reduces p-STAT3 the expression,the expression rate by 58.95% drops is 20.67%, reduced the survivin expression, the expression rate is indirectly 16.24% differences has statistics significance by 56.91% drops (P<0.05). Indicated that survivin the expression activates along with STAT3 is suppressed, the p-STAT3 expression rate drop drops.Conclusion: In Ovary epithelioma SKOV3 cell line, AG490 through suppresses the JAK enzyme activeness, blocks the STAT signal the activation, reduced the p-STAT3 expression, then reduced its downstream the protein survivivn's expression, the promotion tumour cell apoptosis rate is upgrade, suppresses the tumour cell proliferation . It is the ovary epidermis cancer's suppressor gene.