Tao Qin1, Cui-qing Fan1, Ning Zhu2, Yan Shen1, Mei-hong Chen1. 1. State Key Laboratory of Medical Molecular Biology, Department of Biochemistry and Molecular Biology, Institute of Basic Medical Sciences, CAMS and PUMC, Beijing 100005, China. 2. State Key Laboratory of Medical Molecular Biology, Department of Biochemistry and Molecular Biology, nstitute of Basic Medical Sciences, CAMS and PUMC, Beijing 100005, China.
Abstract
OBJECTIVE: To study the effect of human proteasome subunit Α7(PSMA7)gene silencing by small interfering RNA(siRNA)on human myeloid leukemia cell line K562. METHODS: PSMA7 gene-specific siRNA was chemically synthesized and transfected into K562 cell line by HiPerFect. The expression level of PSMA7 protein was detected by Western blot analysis. Cell proliferation was determined by MTS and cell counting. Cell cycle distribution was measured by flow cytometry. The expressions of Cyclin A, D, and E were detected by Western blot analysis. The apoptotic ratio was determined by flow cytometry. RESULTS: PSMA7 protein was evidently silenced 48 hours after transfection of the PSMA7-specific siRNA into K562 cell line. The proliferation of the cells was markedly inhibited, and the percentage of S phase cells decreased. However, no apoptosis was observed. The expressions of Cyclin A and E were down-regulated. CONCLUSION: Knockdown of PSMA7 down-regulates the expression of Cyclin A and E and thus decreases the proportion of cells in S phase as a result, the proliferation of K562 cell line is inhibited.
OBJECTIVE: To study the effect of human proteasome subunit Α7(PSMA7)gene silencing by small interfering RNA(siRNA)on humanmyeloid leukemia cell line K562. METHODS:PSMA7 gene-specific siRNA was chemically synthesized and transfected into K562 cell line by HiPerFect. The expression level of PSMA7 protein was detected by Western blot analysis. Cell proliferation was determined by MTS and cell counting. Cell cycle distribution was measured by flow cytometry. The expressions of Cyclin A, D, and E were detected by Western blot analysis. The apoptotic ratio was determined by flow cytometry. RESULTS:PSMA7 protein was evidently silenced 48 hours after transfection of the PSMA7-specific siRNA into K562 cell line. The proliferation of the cells was markedly inhibited, and the percentage of S phase cells decreased. However, no apoptosis was observed. The expressions of Cyclin A and E were down-regulated. CONCLUSION: Knockdown of PSMA7 down-regulates the expression of Cyclin A and E and thus decreases the proportion of cells in S phase as a result, the proliferation of K562 cell line is inhibited.