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. 2020 Sep 11;10(24):11144–11158. doi: 10.7150/thno.42578

Figure 2.

Figure 2

Cyclin K knockdown suppresses tumorigenesis in lung cancer both in vitro and in vivo. (A) Cyclin K was successfully knocked down by two different siRNAs. A549 (left) or H460 (right) cells were transfected with the indicated siRNAs for 48 h, then collected and analyzed by Western blotting (n = 3). (B) Cyclin K depletion decreased cell growth. A549 and H460 cells were transfected with indicated siRNAs for 48 h and then seeded. Cell numbers were calculated every two days (n = 3). (C) Colony formation was significantly decreased in Cyclin K-depleted cells. A549 and H460 cells transfected with indicated siRNAs were grown for two weeks. Colonies were counted and are shown as histograms (n= 3). *P < 0.05, **P < 0.01, ***P < 0.001 compared to control cells. (D) Cells were transfected with indicated siRNAs for 48 h, then dyed with CFSE stain. Cells were harvested in trypsin at indicated time points and analyzed by flow cytometry (n = 3). (E) H460 cells stably transfected with shCyclin K or shControl were lysed and analyzed by Western blotting (n = 3). (F) Cyclin K knockdown alleviated tumor growth in vivo. Representative images of xenograft tumors are shown (five mice/group). (G) Representative immunohistochemistry images showing Cyclin K and Ki-67 expression in xenograft tumors in the three groups. Scale bar, 50 µm. (H) Growth curves of tumors in the three groups are presented (five mice/group). Data are shown as the mean tumor volume ± SEM. (I) Tumor weight in the three groups are shown (five mice/group).