Aim: To elucidate the effect of p27(KIP1) on cell cycle and apoptosis regulation in gastric carcinoma cells.
Methods: The whole length of p27(KIP1) cDNA was transfected into human gastric cancer cell line SCG7901 by lipofectamine. Expression of p27(KIP1) protein or mRNA was analyzed by Western blot and RNA dot blotting, respectively. Effect of p27(KIP1) on cell growth was observed by MTT assay and anchorage-independent growth in soft agar. Tumorigenicity in nude mice was used to assess the in vivo biological effect of p27(KIP1). Flow cytometry, TUNEL, and electron microscopy were used to assess the effect of p27(KIP1) on cell cycle and apoptosis.
Results: Expression of p27(KIP1) protein or mRNA increased evidently in SCG7901 cells transfected with p27(KIP1). The cell growth was reduced by 31% at 48 h after induction with zinc determined by cell viability assay. The alteration of cell malignant phenotype was evidently indicated by the loss of anchorage-independent growth ability in soft agar. The tumorigenicity in nude mice was reduced evidently (0.55+/-0.14 cm vs 1.36+/-0.13 cm, P<0.01). p27(KIP1) overexpression caused cell arrest with 36% increase (from 33.7% to 69.3%, P<0.01) in G1 population. Prolonged p27(KIP1) expression induced apoptotic cell death reflected by pre-G1 peak in the histogram of FACS, which was also confirmed by TUNEL assay and electron microscopy.
Conclusion: p27(KIP1) can prolong cell cycle in G1 phase and lead to apoptosis. p27(KIP1) may be a good candidate for cancer gene therapy.