The expression of pol ι is dependent on activated JNK/c-Jun.

<p>(A) The kinetics of pol ι, p-JNK and p-c-Jun expression under UV damage in HEK293 cells. (B) The effect of UV damage on the activity of the <i>POLI</i> promoter region. *Statistically significant difference compared to the pGL3-275/+63 and pGL3–275/+63MU construct (p<0.01; Student's t-test). **Statistically significant difference compared to the cells treated with or without UV (p<0.01). (C) The expression of pol ι, c-Jun and p-c-Jun in RT4 and T24 bladder cancer cells. The expression of pol ι was observed in (D) HEK293, T24 and RT4 cells treated with different concentrations of SP600125 cultured for additional different lengths of time; (E) HEK293 cells treated with PD98059 or SB203580; (F) HEK293 cells treated with or without SP600125 and UV; (G) HEK293 cells transfected with siRNA-c-Jun for 48 hours. The non-specific siRNA was used for control. Densitometric values were indicated below each panel.</p

Mutation spectra generated by UVC (1000 J/m²) of <i>SupF</i> gene in T24 cells.

<p>(A) Mutations obtained from the T24 cells (total 60 point mutants in 25 bacterium samples). (B) Mutations obtained from the T24 cells pre-treated with SP600125 (total 55 point mutants in 25 bacterium samples). T24 cells treated by SP600125 two times each for 2 hours (one time every 24 hours for 48 hours) were lysed at 24 hours and 48 hours, and pol ι expression was analyzed by Western blot assay. Densitometric values were indicated below each panel. Point mutations were indicated under each base pair and the listed base represented a change from the top strand. <i>SupF</i> gene sequence was list in double strand.</p

The expressions of pol ι and p-c-Jun in human urothelial carcinoma samples.

<p>(A) pol ι protein is aberrantly expressed in different tumor grade according to the WHO classification. A1 is I-grade tumor; A2 is II-grade tumor; A3 is III-grade tumor (×400 magnification). (B) The expression of pol ι in 5 pairs of bladder carcinoma sides and cancer tissues. “N” was represented carcinoma sides tissue, “C” was represented cancer tissue. Densitometric values were indicated below each panel. (C) Representative cases of the concordance of pol ι and p-c-Jun expression in bladder cancer. C1 and C2 were respectively showed the high expression of pol ι and p-c-Jun in a pairs of simple. C3 and C4 were the low expression in another pairs (×100 and ×400 magnification).</p

The expression of p-c-Jun was correlated with pol ι in bladder cancers.

<p>There are 41 paraffin-embedded bladder tumor tissues from a total of 97 patient samples. The expression of pol ι coincided with p-c-Jun was 34 pairs of tissues. “L/L” was represented both low expression of p-c-Jun and pol ι; “I/I” was represented both high expression of p-c-Jun and pol ι. The number of samples was listed. The concordance of the p-c-Jun and pol ι expression was evaluated by bivariate test; p<0.01, r = 0.662.</p

The expression of pol ι is associated with malignancy in human bladder cancers.

<p>pol ι expression were examined in 97 bladder tumor tissues. “Low (L; %)” or “Intense (I; %)” was respectively represented the number and the percentage in the grade of low or high expression of pol ι. The number of samples and the percentage were listed. Statistically significant difference was analyzed by chi-square test to compare to the different pathological grade of bladder tumor tissues (p<0.01).</p

c-Jun activates and binds to the human <i>POLI</i> promoter.

<p>(A) Luciferase activity of the deletions of 5′ flanking region of <i>POLI</i> gene was normalized by β-<i>gal</i> activity. Each bar represents the mean±SD for at least three independent experiments. (B) Schematic representations of the distal <i>POLI</i> promoter region. (C) Transcriptional activities of the deletion or mutant of <i>POLI</i> promoter. *Statistically significant difference compared to the pGL3-275/+63 and pGL3–275/+63MU construct (p<0.01; Student's t-test). (D) EMSA Assay. (E) ChIP assay. Mouse IgG as a negative control. The Input DNA or no DNA added was each used as positive and blank control, respectively.</p

Mutation frequency of <i>SupF</i> gene in T24 cells.

*<p>Each data at least three independent experiments. The mutation frequency (×10⁻³) was described by mean± SE (SE: standard error).</p

Inhibition of ATR or XPC reduced UV-induced pol ι expression and abolished JNK/c-Jun activation.

<p>Protein expression was investigated in different cells with different treatment. (A) HEK293 cells transfected with siRNA-ATR. (B) HEK293 cells infected with shRNA-XPC. (C) T24 cells transfected with siRNA-ATR. (D) T24 cells infected with shRNA-XPC. (E) Real-time PCR analysis of the expression of MKP-1 in untreated or treated HEK293 cells with siRNA-c-Jun, siRNA-ATR or shRNA-XPC; GAPDH mRNA levels were determined as an internal control. *Statistically significant difference compared to HEK293 cells with or without treatment (p<0.01; Student's t-test). (F) MKP-1 protein expression in HEK293 cells treated with siRNA-ATR or shRNA-XPC. The non-specific siRNA or non-specific shRNA-lentivirus was used for control. Densitometric values were indicated below each panel.</p

Primers used in PCR for <i>POLI</i> gene.

<p>Primers used in PCR for <i>POLI</i> gene.</p

Hyaluronic Acid-Conjugated Fluorescent Probe-Shielded Polydopamine Nanomedicines for Targeted Imaging and Chemotherapy of Bladder Cancer

Bladder cancer is one of the most common malignancies in the urinary system, with high risk of recurrence and progression. However, the difficulty in detecting small tumor lesions and the lack of selectivity of intravesical treatment seriously affect the prognosis of patients with bladder cancer. In the present work, a nanoparticle-based delivery system with tumor targeting, high biocompatibility, simple preparation, and the ability to synergize imaging and therapy was fabricated. Specifically, this nanosystem consisted of the core of doxorubicin (DOX)-loaded polydopamine nanoparticles (PDD NPs) and the shell of hyaluronic acid (HA)-conjugated IR780 (HA-IR780). The HA-IR780-covered PDD NPs (HR-PDD NPs) demonstrated tumor targeting and visualization both in vitro and in vivo with properties of promoted cancer cell endocytosis and lysosomal escape, efficiently delivering drugs to the target site and exerting a killing effect on tumor cells. Encouragingly, intravesical instillation of HR-PDD NPs improved drug retention in the bladder and promoted its accumulation in tumor tissue, resulting in better tumor proliferation inhibition and apoptosis in an orthotopic bladder cancer model in rats. This study provides a promising strategy for the diagnosis and therapy of bladder cancer