Fig. 5.

(a) A colony formation experiment was performed to detect the effect of RGD4C-scFv on SW480 cell proliferation. SW480 cells were incubated with 20 μM fusion protein. After 2 weeks of incubation, monoclonal cells were stained with Giemsa. The numbers of tumor cell clones in the RGD4C-scFv and RGD4C-linker-scFv groups were significantly lower than those in the anti-p21Ras scFv and PBS groups. (b) The clone numbers of SW480 cell in the RGD4C-scFv and RGD4C-scFv+CQ groups were also significantly lower than those in the RGD4C-EGFP and PBS groups. (c) However, CACO-2 cell clones had no significant difference between the experimental group and the control group. (d) The clone numbers of normal cell CCD841 cells in the RGD4C-scFv and RGD4C-scFv+CQ groups were roughly the same with those in the RGD4C-EGFP and PBS groups. (e) After treatment with RGD4C-p21Ras-scFv for 1d, 2 d, or 3 d, the proliferative activity of SW480 cells was tested by an MTT assay. The growth of SW480 cells was inhibited by both RGD4C-scFv and RGD4C-linker-scFv compared with the anti-p21Ras scFv and PBS. (f) After treatmented with RGD4C-scFv, RGD4C-EGFP or RGD4C-scFv+CQ for 1d, 2 d, 3 d, the growth of SW480 cells was inhibited by both RGD4C-scFv or RGD4C-scFv+CQ compared with the RGD4C-EGFP and PBS. (g and h) After treatment with RGD4C-scFv, RGD4C-EGFP or RGD4C-scFv+CQ for 1d, 2 d, 3 d, neither the RGD4C-EGFP and PBS control groups nor the RGD4C-scFv and RGD4C-scFv+CQ experimental group had any killing effect on the CACO-2 and CCD841 cells