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Ectopic expression of guanylyl cyclase C in gastric cancer as a potential biomarker and therapeutic target
Author(s) -
MAO Zhen Biao,
ZHANG Jian Feng,
XU Zhong,
ZHU Hui Jun,
ZHANG Jian Guo,
PAN Zheng Ping,
XIAO Feng,
YANG Jun Ling
Publication year - 2009
Publication title -
journal of digestive diseases
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.684
H-Index - 51
eISSN - 1751-2980
pISSN - 1751-2972
DOI - 10.1111/j.1751-2980.2009.00396.x
Subject(s) - microbiology and biotechnology , cell cycle , apoptosis , transfection , cell growth , small interfering rna , flow cytometry , viability assay , cell , gene silencing , cell culture , biology , gene , biochemistry , genetics
OBJECTIVE: To investigate the expression of guanylyl cyclase C (GCC) in human gastric cancer (GC) tissues and assess the effect of GCC small interfering RNA (siRNA) on the proliferation and apoptosis of SGC‐7901. METHODS: The expression of GCC in 30 specimens and three human GC cell lines (SGC‐7901, AGS, NCI‐N87) were detected by RT‐PCR for messenger RNA (mRNA) by Western blot and immunofluorescence for proteins. Recombinant plasmids containing GCC siRNA and scrambled siRNA were constructed and transfected into SGC‐7901 cells, respectively. A cell counting kit‐8, flow cytometry (FCM) and terminal deoxynucleotidyl transferase (TDT)‐mediated dUTP‐biotin nick end‐labeling were used to evaluate cell viability, cell cycle distribution and apoptosis, followed by wound healing assay and cell adherent assay for cell motility and adherent, respectively. RESULTS: The expression of GCC was absent in paracancerous tissues, whereas the GCC mRNA and protein expressions were detected in 20/30 and 19/30 of GC specimens, respectively. Moreover, intestinal GC was statistically different from diffuse GC ( P < 0.05). The proliferation of SGC‐7901 cells was markedly inhibited by GCC siRNA‐3 ( P < 0.05) and cell morphological changes including volumetric reduction, karyopyknosis and karyorrhexis were observed. FCM showed that the cell count in the sub‐G0/G1 peak increased from 5.47% (48 h after transfection) to 5.63% (72 h after transfection). The wound healing assay and cell adherent assay revealed that GCC gene silencing decreased cell motility and adherent. CONCLUSION: The over‐expression of GCC has been detected in intestinal type GC. GCC siRNA can effectively inhibit the proliferation and invasion of SGC‐7901 cells and induce cell apoptosis. GCC might be a novel biomarker and therapeutic target for GC.