ABSTRACT
Objective: To observe the effects of follistatin (FST)on the skeletal muscle wasting of cancer cachexia mice and the expressions of Mstn, LncRNA-MALAT1 and Caspase-3, and to elucidate its associated molecular mechanisms.Methods:Thirty-two BALB/c mices were randomly assigned into:healthy control (HC) group,FST prevention (FP)group,FST treatment (FT)group and cancer cachexia (CC)group.The murine colon adenocarcinoma CT26 cells were inoculated subcutaneously into the mices in FP, FT and CC groups to establish the cancer cachexia models. The body weight, spontaneous activity and tumor growth were daily monitored.The mice in FP and FT groups were administrated with FST intraperitoneally on day 6 and 12 after inoculation.The samples were collected on day 20.The tumor and gastrocnemius weights of the mice were detected. The biochemical metabolism indexes and myofiber cross-sectional area of gastrocnemius tissue were detected.The mRNA expression levels of Mstn,Caspase-3 and LncRNA-MALAT1 were examined by Real-time PCR.The protein expression levels of Mstn and Caspase-3 were measured by Western blotting method. Results:Compared with CC group,the body weights,spontaneous activities,gastrocnemius weights and myofiber cross-sectional areas were increased (P <0.05);the serum levels of glucose,total protein and albumin of the mice in FP and FT groups were increased (P <0.05).The protein and mRNA expression levels of Mstn and Caspase-3 in gastrocnemius of the mice in CC group were significantly higher and the expression level of LncRNA-MALAT1 was significantly lower than those in HC group (P < 0.05).The mRNA and protein expression levels of Mstn and Caspase-3 in FP and FT groups were reduced and the expression level of LncRNA-MALAT1 was increased compared with CC group (P < 0.05).The prevention effect in FP group is better than FT group (P < 0.05). Conclusion:FST may alleviate the muscle wasting of the mice with cancer cachexia by inhibiting the expression of Mstn,thus upregulating the expression of LncRNA-MALAT1 which in turn to suppress the expression of Caspase-3.
ABSTRACT
Objective To explore the influence of tranSCription factor FOXC2 in angiogenesis of breast cancer MCF-7 cells and to clarify the action mechanism of FOXC2 in promoting tumor angiogenesis.Methods FOXC2 gene and empty vector gene were transfected into breast cancer of MCF-7 cell line with FOXC2 lentivirus gene transfection technique to obtain stable transfection cell line. The MCF-7 cells were devided into non-transfected group,empty-vector group and over-expression group.Matrigel assay and Transwell chamber test were used to observe the changes of tube formation and migration ability of human umbilical vein endothelial cells (HUVECs)in MCF-7 cells supernatant in various groups. PT-PCR and Western blotting methods were applied to detect the expressions of FOXC2,DLL4 and Notch1 mRNA and protein.Results Compared with non-tranfected group and empty-vector group,the tube formation and the migration number of HUVECs in FOXC2 over-expression group were increased(P<0.05);the expressions of FOXC2,DLL4 and Notch1 mRNA and proteins were significantly increased(P<0.05).Conclusion The FOXC2 over-expression in MCF-7 cells can increase the tube formation ability and migration ability of HUVECs,and its mechanism may be related to Notch signaling pathway.
ABSTRACT
Objective To explore the expression of caspase-3 in skeletal muscle of the mice in the state of cancer, and to elucidate the relationship between Caspase-3 and apoptosis,consumption of skeletal muscle protein in cancer cachexia.Methods 48 male BALB/c mice were randomly divided into cancer cachexia group and control group (n=24).The mice in cancer cachexia group were inoculated with mouse colon 26 adenocarcinoma cells.The body weights of the mice in two groups were detected daily.Eight mice in each group were executed to test the weight of left gastrocnemius, fiber crosscut area, the expression levels of tumor necrosis factor-α(TNF-α), interleukin-6 (IL-6),Caspase-3 proteins and the apoptotic rate of gastrocnemius cells on day 8,14,and 20,respectively. Results The mice in cancer cachexia group appeared cachectic symptoms on day 14,the non-tumor body weight was decreased more than 20% of that in control group (P<0.05).Compared with control group at the same time, the mouse body weight,non-tumor body weight,the weight of left gastrocnemius and the fiber crosscut area of the mice in cancer cachexia group were obviously decreased with the prolongation of inoculation time (P<0.05 ), whereas the expression levels of TNF-α,IL-6,Caspase-3 proteins and the apoptotic rate of muscle cells were obviously increased after tumor inoculation (P<0.05).The level of Caspase-3 protein was negatively correlated with the weight of gastrocnemius and fiber crosscut area (r=-0.716,P<0.05;r=-0.694,P<0.05),and the level of Caspase-3 was positively correlated with the levels of TNF-αand IL-6 (r=0.742,P<0.05;r=0.675,P<0.05).Conclusion Caspase-3 may be a key factor in the protein comsumption of skeletal muscle in cancer cachexia.