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. Author manuscript; available in PMC: 2010 Jun 1.
Published in final edited form as: Lung Cancer. 2009 Mar 28;66(3):292–297. doi: 10.1016/j.lungcan.2009.02.014

Figure 1.

Figure 1

Expression of KLF6-SV1 in cisplatin resistant lung cancer cell lines. a, Generation and validation of cisplatin resistance in the A549 lung adenocarcinoma cell line. The A549-IR and A549-CR cells were generated by serial selection of the parental A549 cell line in 20 μM cisplatin for 2 and 8 passages respectively. All cell lines were plated at equal densities and treated with 20uM of cisplatin, 72 hrs after treatment cells were harvested. A, Representative FACS analysis of the A549-NR, A549-IR and A549-CR cell line treated with 20 μM cisplatin. FACS analysis of the treated cell lines revealed a marked reduction in the induction of apoptosis in parental A549 cell line (A549-NR) (33.7%) vs. A549-IR (16.7%) vs. A549-CR (4.4%) cell lines. A marked induction of apoptosis is seen in the control A549 cell line while addition of cisplatin results in a G2/M arrest with little induction of apoptosis in the A549-CR cell line. The A549-IR cell line displayed an intermediate sensitivity to cisplatin. b, A significant decrease in apoptosis in the A549-IR and A549-CR compared to the control A549 cell line (*** p < 0.001). This was repeated three independent times. C, qtRT-PCR and western blot analysis of the A549-NR, A549-IR, and A549-CR demonstrate a marked upregulation of KLF6-SV1 at both the mRNA and protein level (** p < 0.001). d, p21 expression at both the mRNA and protein level in the A549-CR cell line is significantly lower than the parental A549-NR cell line (** p < 0.001). e, the ration of KLF6-SV1 to wtKLF6 was determined by quantitative real time PCR (qRT-PCR) using KLF6-SV1 and wtKLF6 specific primers. There is a significant increase in the ratio of KLF6-SV1/wtKLF6 in the cisplatin resistant cell line.