For in vivo metastasis assays, each experimental group consisted of six 6-week-old male BALB/c-nu/nu mice. Briefly, 1.5 × 107 cells with stably COL6A1 overexpressed or knockdown were suspended in 40 μL serum-free DMEM for each nude mouse (6 in each group), were injected intravenously through the tail vein into mouse. The mice were killed 8 weeks after injection. Tumor nodules formed on the lung surfaces were macroscopically determined and counted. The lungs were excised and embedded in paraffin. Our protocol was approved by the Committee on the Ethics of Animal Experiments of Sun Yat-sen University Cancer Center.

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