Affiliations: Qingpu Branch, Affiliated Zhongshan Hospital, Fudan University, Shanghai, China
Correspondence:
[*]
Corresponding author: Guang-Ping Bai, Park East Road 1158, Qingpu District, Shanghai 201700, China. Tel./Fax: +86 21 69719190; E-mail:gping_bai@126.com
Abstract:
AIMS:
To clone and express Siva1 protein, and to
investigate the role of Siva1 protein in proliferation, apoptosis, invasion,
and migration of human nasopharyngeal carcinoma cell line CNE-2 in vitro
and in vivo. METHODS:
The PCR fragment of Siva1 from human nasopharyngeal carcinoma cell
line CNE-2 were double digested with BamHI and SalI and then induced into
the pQE30 vector double digested by the same enzymes. The pQE30 vector
harboring Siva1 was introduced into M15 competent cells and then induced by
isopropyl β -D-1-thiogalactopyranoside (IPTG). The Siva1 fusion
protein was identified by 12% sodium dodecyl sulfate polyacrylamide gel
electrophoresis (SDS-PAGE), and then separated and purified by Ni-affinity
chromatography. Subsequently, the effects of recombinant Siva1 protein on
proliferation, apoptosis, invasion and migration were assayed in vitro and in vivo. RESULTS: The transformed cells expressed Siva1 fusion protein with
a molecular weight of approximately 12 kDa. Cell counting kit-8 (CCK-8)
assay showed that the Siva1 protein significantly inhibited the
proliferation of the CNE-2 cells at a concentration of 10 μ mol/L. In
addition, compared to the control, the Siva1 protein promoted the apoptosis
of the cancer cells. And, the Siva1 protein greatly suppressed the invasion
and migration of the cancer cells. In vivo, the Siva1 protein significantly
inhibited the tumor growth of the tumor-bearing mice. Further, the Siva1
treatment markedly upregulated Bax, caspase-3, and downregulated Bcl-2
protein levels in the transplanted tumor tissue. CONCLUSIONS: The
Siva1 protein has a significant anticancer activity on human nasopharyngeal
carcinoma cell line CNE-2 including inhibiting proliferation, invasion,
migration and promoting apoptosis of the cancer cells.
