Abstract: Objective To establish a OVCAR3 cell line stably expressing inducible shRNA targeting CKS2 and explore the effects of CKS2 knockdown on cell proliferation of OVCAR3. Methods Two oligonucleotides targeting CKS2 gene were synthesized and cloned into lentivirus expression plasmid pLVTHM, thus, pLVTHM/CKS2 shRNA recombinant plasmids were constructed. OVCAR3 cells were firstly infected with lentivirus made from pLV-tTR/KRAB-Red empty vector, followed by infecting with another kind of lentivirus prepared from pLVTHM/CKS2 shRNA recombinant plasmid. The expression of CKS2 mRNA and protein were determined by real time PCR and Western blot in OVCAR3 cells infected with lentivirus and treated with doxycycline (DOX) for 72h, respectively. The effects of cell proliferation were determined by MTT. Results The lentivirus expression plasmids containing CKS2 shRNA were constructed successfully, the lentivirus were produced and OVCAR3 cells were infected by these lentivirus. A OVCAR3 cell line stably expressing inducible shRNA targeting CKS2 was established successfully and the CKS2 expression levels were decreased obviously after induction by DOX（P < 0.05，P < 0.01）. CKS2 knockdown could inhibit OVCAR3 cell proliferation. Conclusions A OVCAR3 cell line stably expressing inducible shRNA targeting CKS2 was established successfully, CKS2 expression suppression induced by DOX could inhibit OVCAR3 cell proliferation.

Key words: CKS2, Inducible shRNA, Ovarian cancer