Effects Of Hedgehog Signaling Pathway On Proliferation And Apoptosis Of Colorectal Cancer Cells

Objective:1. To research the effects of blockage of hedgehog signaling on proliferation,apoptosis of colorectal cancer cell sw480and caco2.2. To research the distinction of the activating mechanism on hedgehog signaling between sw480and caco2.Methods:1. cultivating sw480 and caco2 cell lines, observating cell morphous under inverted microscope.2.cells were treated with Cyclopamine:(1) observating cell morphous diversify, (2) cell proliferation was detected by MTT, (3) cell apoptosis was analysed by flow cytometry.3 constructing Glil miRNA interference plasmid which were transfected into sw480 cell, Gli1 expression in sw480 cell was assayed by by RT-PCR.. cell proliferation was detected by MTT.Results:1. after cyclopamine treatment:(1) intercellular space of caco2 was larger, cell morphous changeed significantly in a dose-and-time dependent manner; cell morphous of SW480 had not changed significantly. (2)cyclopamine could significantly inhibit the proliferation of caco2s in a dose-and-time dependent manner.40μmol/L,72h. inhibition ratio of proliferation reached up to 83.69%. but to SW480s, cell proliferation could not be inhibited significantly (p>0.05).(3) compared with control group (14.44%),treatment of caco2s with cyclopamine induced a remarkable increase of apoptosis ratio.40μmol/L,20μmol/L,10μmol/L,48h. apoptosis ratio respectively reached up to 64.03%,50.47%,31.7%(P<0.05)。2. After 48 hours of transfecting Glil miRNA interference plasmid into sw480 cell, (1)Gli1 mRNA in sw480 cell decreasd up to80%; (2)intercellular space of Sw480 became larger; (3)the proliferation of sw480s could be significantly inhibited, inhibition ratio of proliferation reached up to 37.44%。Conclusion:blockage of hedgehog signaling could inhibit the cell proliferation both in caco2andSW480; the proliferation of caco2 and SW480 were controlled by hedgehog, but the route of activating the hedgehog signaling is different between caco2andSW480.to caco2,hedgehog was activated by classical pathway,to SW480, hedgehog was activated by alternative pathway.