Fig 1: SPARC is associated with prognosis of various cancers. (A-I) High SPARC expression is associated with worse prognosis in ACC, GBM, KIRP, LUSC, MESO, PAAD, SARC, STAD and UVM (TGCA data). (J-M) High SPARC expression is associated with improved prognosis in DLBL, KIRC, LGG and SKCM. (TCGA data). The Kaplan-Meier method with log-rank tests (ACC, GBM, LUSC, MESO, PAAD, SARC, STAD, UVM, DLBL, KIRC) or Cramér-von Mises tests (KIRP, LGG, SKCM) were used for comparison of survival time between two groups. SPARC, secreted protein acidic and rich in cysteine; ACC, adrenocortical carcinoma; GBM, glioblastoma multiforme; KIRP, kidney renal papillary cell carcinoma; LUSC, lung squamous carcinoma; MESO, pleura mesothelioma; SARC, soft tissue sarcoma; STAD, stomach adenocarcinoma; UVM, uveal melanoma; DLBL, diffuse large B-cell lymphoma; KIRC, kidney renal clear cell carcinoma; LGG, brain low-grade glioma; SKCM, skin cutaneous melanoma.
Fig 2: SPARC drives tumorigenesis through abnormal changes in expression but not gene mutations. (A) Single-nucleotide variations are uniformly distributed across the coding region of the SPARC gene (TCGA data). (B) Expression pattern of SPARC in 33 cancer types. SPARC expression is generally increased in various cancer types. Unpaired, two-tailed t-test (TCGA data). (C) Expression of SPARC decreased significantly in recurrent LGG, compared with the primary tumour. Unpaired, two-tailed t-test (TCGA data). (D) Expression of SPARC decreased significantly in recurrent OV, compared with the primary tumour. Unpaired, two-tailed t-test (TCGA data). (E) Reverse transcription-quantitative PCR demonstrated that SPARC expression increased significantly in PAAD, compared with the paired para-cancer tissue. n=17, paired, two-tailed t-test. *P<0.05. LGG, brain low-grade glioma; OV, ovarian serous cystadenocarcinoma; PAAD, pancreatic adenocarcinoma; TCGA, The Cancer Genome Atlas; FPKM, fragments per kilobase million; SPARC, secreted protein acidic and rich in cysteine.
Fig 3: The expression of proteins of these common differential genes in tissuesThe expression levels of protein COL12A1 (G), COL1A2 (D), COL4A1 (E), SPARC (K), COL5A2 (F) and COL1A1 (C) was increased, while ACAA1 (A), ACOX1 (B), CPT2 (H), SQRDL (L), HMGCS2 (J), ETHE1 (I) and TST (M) were decreased. *P < 0.05.
Fig 4: The expression of mRNA of these common differential genes in tissuesThe expression levels of mRNA COL12A1 (G), COL1A2 (D), COL4A1 (E), SPARC (K), COL5A2 (F) and COL1A1 (C) was increased, while ACAA1 (A), ACOX1 (B), CPT2 (H), SQRDL (L), HMGCS2 (J), ETHE1 (I) and TST (M) were decreased. *P < 0.05.
Fig 5: SPARC regulates pancreatic cancer cell proliferation through autocrine secretion into the extracellular milieu. (A) SPARC is detected in cell lysate and supernatant of PAAD cells, immortalized normal acinar AR42Jccells, and 293T cells. (B) Supernatant of 293T cells transfected with the SPARC siRNA has significant decreased concentration of SPARC protein, compared with cells transfected with scramble siRNA. The supernatant of 293T cells transfected with SPARC OE vector has increased concentration of SPARC protein, compared with the empty vector group. (C) Proliferation of PANC-1 cells transfected with SPARC OE supernatant increased significantly, compared with the cells treated with the SPARC-silenced supernatant (n=3). *P<0.05, unpaired two-tailed t-test. (D) Proliferation of SW1990 cells treated with the SPARC OE supernatant increased significantly, compared with the cells treated with the SPARC-silenced supernatant (n=3). *P<0.05, unpaired two-tailed t-test. (E) Treatment with SPARC OE supernatant restores clone formation in PANC-1 and SW1990 cells. (F) Schematic plot of the construction of SPARC∆sig peptide-Flag expression vector. (G) SPARC∆sig peptide-Flag is expressed but not effectively secreted to the extracellular milieu. (H and I) Wild-type SPARC, but not SPARC∆sig peptide, promotes the proliferation of (H) PANC-1 cells and (I) SW1990 cells (n=3). *P<0.05, one-way ANOVA with Tukey's post-hoc test. (J) Colony formation in PAAD cells is restored by wild-type SPARC, but not SPARC∆sig peptide. Quantitative data from three independent experiments was presented as mean ± SD (error bars). OE, overexpression; SPARC, secreted protein acidic and rich in cysteine; siRNA, small interfering RNA.
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