Inhibitions Of SAHA Combined With DDP On Cervical Cancer In Nude Mice Xenografts

Objective Through observe the functions of SAHA in two doses or combined withcisplatin to cervical cancer transplanted in nude mice, detect the related VEGF and Baxprotein expressions changes, discuss SAHA and it combined cisplatin in cervical cancerin nude mice and its possible mechanism, to provide experimental basis for clinical.Method SiHa cells cultivated and inoculated into nude mice to establish nude mousecervical cancer transplantation tumor model.6groups randomly assigned into after tumorestablished succeed,and each group has6nude mice:1）blank control group:normalsaline2）DDP group：DDP5mg/kg3）SAHA1group：SAHA25mg/kg4）SAHA2group：SAHA50mg/kg5）Joint group1：SAHA25mg/kg+DDP5mg/kg6）Jointgroup2：SAHA50mg/kg+DDP5mg/kg. Intraperitoneal injection, for eight times:oncea day for four days, and then consecutive four days after intermittent seven days. Recordtumor growth, general conditions and body weight of the nude mice, measure the lengthof the tumors every other day in intervention period, calculate the volume of tumors anddraw the growth curve. Killed the mude mice after withdrawal two days,peel andweight the tumor, calculate the tumor growth inhibition rate. HE staining to observe thetumor pathological changes. Immunohistochemistry and Western blot assay thetransplanted tumors Bax, VEGF factor expression changes.Result1The establishment of animal model end general conditions of nude mice inexperiment period There are36nude mice were transplanted successed in40nude miceinoculated SiHa cells，the tumor formation rate was90%. After the interventioncompared with before the intervention, the body weight of nude mice in blank controlgroup and SAHA1group increased (P<0.05), where as in the DDP group and jointgroups decreased (P<0.05).After the intervention, the body weight of nude mice hasstatistically significant (F=92.978,P<0.05), the body weight of nude mice in interventiongroup were less than in blank control group, nude mice in joint group2and DDP grouphas the least body weight (P <0.05), and during intervention,the nude mice with slowactivity,in depression, whose appetite declined gradually and subcutaneous fatdisappeared gradually, got skin without luster and stool thin, etc. There was a nude micedied after12days intervented.2transplanted tumors volume change and transplantedtumor weight inhibition rate Intervene started after inoculation after3weeks,transplanted tumors volume of nude mice increased gradually. After the intervene, thetransplanted tumors volume of nude mice in experimental group were increasedcompared with before the intervene(P<0.05). After the intervene, the transplanted tumorsvolume of nude mice in intervention group were less than in blank control group(P<0.05), and transplanted tumors volume of SAHA2group was less than SAHA1group(P<0.05), the transplantated tumors volumes of nude mouse in joint groups were less than each single drug intervention group(P<0.05), but there is no statistically significantdifference with transplanted tumors volume between in joint group2and joint group1(P>0.05). Killed the nude mice after the intervention ended,peel and weight the tumor,calculate the tumor inhibition rate: blank control group0, DDP group43.64%,SHAH1group20.33%,SAHA2group32.73%,joint group155.90%, joint group256.30%. The Q value of25mg/kg SAHA combined with5mg/kg DDP was1.015,The Qvalue of50mg/kg SAHA combined with5mg/kg DDP was0.907.3xenograft tumorpathology observed Transplanted tumors were solid or semi-solid, clear, fish-shapedcut surface boundaries. In HE staining microscope, cells in intervention groups has bodylager, vacuoles degenerated,periplasm looser, ucleus structure disappeared, and somecells has ucleus pyknotic cracking. necrotic cells and tissue were visible, the mostobvious phenomenon were in the joint groups.4Immunohistochemical detection of eachgroup Bax, VEGF positive expression Bax positive expression locate in cytoplasm,compared with in the control group and SAHA1group, the Bax positive staining index inother groups were significantly higher (P <0.05), the the Bax positive staining index injoint groups were the highest, but there was no statistical differences between in the twojoint groups (P>0.05). VEGF positive expression locate in cytoplasm, compared with inthe control group and SAHA1group, the VEGF positive staining index in other groupswere significantly lower (P <0.05), the the VEGF positive staining index in joint groupswere the lowest, but there was no statistical differences between in the two joint groups(P>0.05).5Bax protein expression of transplanted tumor tissue in each groups: in theintervention groups,it is higher than in blank control group (P<0.05); in SAHA2group itis higher than in SAHA1group (P<0.05); in the two joint groups it is higher than insingle intervention groups,where as there has no difference between in the two jointgroups (P>0.05). VEGF protein expression of transplanted tumor tissue in each groups:in the intervention groups,it is lower than in blank control group (P<0.05); in SAHA2group it is lower than in SAHA1group (P<0.05); in the two joint groups it is lower thanin single intervention groups,where as there has no difference between in the two jointgroups (P>0.05).Conclusion1SAHA at25mg/kg and50mg/kg two concentrations has a inhibitoryeffec to cervical cancer transplanted into nude mice.2SAHA combined with DDP canmore inhibit cervical cancer SiHa cells transplanted into nude mice,and the combinationeffect is a additive effect.3the inhibit effects of SAHA or combined with DDP to SiHacell tranplanted tumor growth may be relevant to the up-regulated Bax expression and thedown-regulated VEGF expression.