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. Author manuscript; available in PMC: 2015 Apr 17.
Published in final edited form as: Cell Rep. 2015 Mar 26;11(1):111–124. doi: 10.1016/j.celrep.2015.03.002

Figure 6. Rab2A and Its Q58H Mutant Endow BCSC Traits to Normal Primary Human MECs, whereas Silencing Rab2A Inhibits Primary Human BCSC Expansion and Tumorigenesis.

Figure 6

(A) Western blot showed lentivirus-mediated overexpression of Rab2A and Q58H mutant in two cases of human normal Lin MECs. Arrowhead, exogenous Flag tagged protein; arrow, endogenous protein.

(B) Rab2A or Rab2A Q58H mutant increased the CD24CD44+ population in primary human MECs.

(C) Real-time PCR showed that expression of Rab2A mRNA was markedly increased in the LinCD24CD44+ population, compared to Linnon-CD24CD44+ or normal epithelial cells.

(D) Expression of Rab2A and unphosphorylated β-catenin protein was markedly increased in LinCD24CD44+ cells compared to Linnon-CD24CD44+ cells in human breast cancer tissue and those in normal breast tissue from the same patient.

(E) Rab2A was knocked down in LinCD24CD44+ cells sorted from human breast cancer tissue.

(F) Rab2A KD in LinCD24CD44+ breast cancer cells decreased the CD24CD44+ population.

(G and H) Rab2A KD in LinCD24CD44+ breast cancer cells decreased mammosphere formation. Scale bar represents 100 μm.

(I–K) Rab2A KD interfered with both tumor initiation and growth of primary BCSCs in vivo, as shown by tumor growth curve (I), tumor weights (J), and tumor incidence (K). 2,000 lentivirus-transduced LinCD24CD44+ cells isolated from eight breast cancer patients were serially transplanted as xenografts into eight nude mice. P0, freshly isolated primary cells; P1, passage 1; P2, passage 2.

In (C) and (H), error bars represent SD of three independent experiments. In (I) and (J), error bars represent SD of eight mice. See also Figure S6.