MicroRNA-205-5p inhibits skin cancer cell proliferation and increase drug sensitivity by targeting TNFAIP8.

Tumor necrosis factor-α-induced protein 8 (TNFAIP8) is a member of the TIPE/TNFAIP8 family which regulates tumor growth and survival. Our goal is to delineate the detailed oncogenic role of TNFAIP8 in skin cancer development and progression. Here we demonstrated that higher expression of TNFAIP8 is associated with basal cell carcinoma (BCC), squamous cell carcinoma (SCC), and melanoma development in patient tissues. Induction of TNFAIP8 expression by TNFα or by ectopic expression of TNFAIP8 in SCC or melanoma cell lines resulted in increased cell growth/proliferation. Conversely, silencing of TNFAIP8 decreased cell survival/cell migration in skin cancer cells. We also showed that miR-205-5p targets the 3'UTR of TNFAIP8 and inhibits TNFAIP8 expression. Moreover, miR-205-5p downregulates TNFAIP8 mediated cellular autophagy, increased sensitivity towards the B-RAFV600E mutant kinase inhibitor vemurafenib, and induced cell apoptosis in melanoma cells. Collectively our data indicate that miR-205-5p acts as a tumor suppressor in skin cancer by targeting TNFAIP8.

[Overexpression of JNK2 enhances proliferation and anti-apoptotic ability of SCL-1 human cutaneous squamous cell carcinoma cells].

Objective To investigate the effect of c-Jun N-terminal kinase 1 (JNK1) or JNK2 overexpression on the proliferation and apoptosis of SCL-1 human cutaneous squamous cell carcinoma cell lines. Methods JNK1 and JNK2 were separately combined with pHBAD-EF1-MCS-3FLAG-CMV-GFP vector to construct the recombinant adenovirus expression vectors of JNK1 and JNK2. The recombinant vectors were used to infect SCL-1 cells. After the optimization of the infection conditions, the fold changes of over-expression were identified by real-time quantitative PCR (qRT-PCR) and Western blot analysis. CCK-8 assay was performed to determine the proliferative activity of SCL-1 cells. Cell colony forming ability was evaluated by cell colony formation assay. Wound healing assay was used to detect the scratch healing rate of SCL-1 cells. Cell apoptosis was determined by flow cytometry. The qRT-PCR was used to detect the mRNA levels of JNK1, JNK2 and c-Jun. The protein level of phosphorylated c-Jun was tested by Western blot analysis. Results The optimal infection condition of JNK1- and JNK2-overexpressing adenoviral vectors was multiplicity of infection (MOI) of 100. The expression levels of JNK1 and JNK2 in SCL-1 cells significantly increased 48 hours after the infection. Compared with the control group, the over-expression of JNK1 had no significant effect on the proliferation and anti-apoptosis ability of SCL-1 cells. The proliferation and anti-apoptosis ability of Ad-JNK2 was significantly enhanced compared with Ad-JNK1, and the phosphorylated c-Jun protein level was up-regulated. Conclusion JNK2 has the function of enhancing the proliferation and anti-apoptotic ability of SCL-1 human cutaneous squamous cell carcinoma cells.

Presence of high risk HPV DNA but indolent transcription of E6/E7 oncogenes in invasive ductal carcinoma of breast.

BACKGROUND AND AIMS: The causative role of high risk human papillomavirus (HR-HPV) in breast cancer development is controversial, though a number of reports have identified HR-HPV DNA in breast cancer specimens. Nevertheless, most studies to date have focused primarily on viral DNA rather than the viral transcription. The aim of this study was to investigate the presence of HR-HPV in breast cancer tissues at HPV DNA level and HPV oncogenes mRNA level by in situ hybridization (ISH). METHODS: One hundred and forty six (146) cases of breast invasive ductal carcinoma(IDC) and 83 cases of benign breast lesions were included in the study. Type specific oligonucleotide probes were used for the DNA detection of HPV 16,18 and 58 by ISH. HR-HPV oncogenes mRNA was assayed by novel RNAscope HR-HPV HR7 assay ISH. p16 protein expression was evaluated by immunohistochemistry (IHC). RESULTS: HR-HPV 16,18 and 58 DNA were detected in 52 out of 146 (35.6%) IDC and in 3 out of 83 (3.6%) benign breast lesions by ISH. The HR-HPV mRNAs was detected only in a few specimens with strong HPV DNA positivity(4/25) in a few scattered cancer cells with very weak punctate nuclear and/or cytoplasmic staining. p16 over-expression did not correlate with the HPV DNA positive breast cancer samples(17/52 HPVDNA+ vs 28/94 HPV DNA-, p=0.731). CONCLUSIONS: HR-HPVs certainly exist in breast cancer tissue with less active transcription, which implies that the causal role of HPV in breast cancer development need further study.

Inhibition of MAPK signaling pathways enhances cell death induced by 5-Aminolevulinic acid-photodynamic therapy in skin squamous carcinoma cells.

Combination of a photosensitizer, 5-aminolevulinic acid (ALA), with photodynamic therapy (PDT) has been widely used to treat skin squamous cell carcinoma (SCC). However, a portion of SCC patients do not respond well to PDT. The molecular reason for this resistance is not clear. We hypothesize that mitogen-activated phosphorylation kinase (MAPK) plays a key role in mediating SCC resistance to PDT. To determine whether inhibition of MAPK signaling enhances the anti-tumor effect of ALA-PDT in SCC. The human squamous carcinoma cell line, SCL-1, was either untreated or treated with various combinations of ALA, PDT light source and inhibitors of MAPK signaling components. ALA-PDT treatment significantly decreased cell viability, increased the percentage of annexin-V positive cells and resulted in formation of apoptotic bodies. ALA-PDT treated cells showed increased levels of p-MEK, p-ERK1/2, p-p38, p-Elk-1, p-JNK and p-c-Jun. Addition of inhibitors for ERK1/2 (PD98059), p38 (SB203580) and JNK (SP60125) reversed the changes and led to a more dramatic decrease in SCL-1 cell viability than seen with ALA-PDT alone. Inhibition of the MAPK pathway enhances the cytotoxic effect of ALA-PDT on SCL-1.

Clinical analysis of five methods used to treat condylomata acuminata.

OBJECTIVE: Treatments for condylomata acuminata (CA) include pharmacotherapy and surgical therapies, but each treatment has its limitations. The aim of this study was to investigate the virus clearance rate, wart cure rate and safety of 5 methods on CA. METHODS: 361 patients diagnosed with CA were divided into groups A (<0.5 cm), B (0.5-2.0 cm) and C (>2.0-4.0 cm) according to the maximum diameter of their lesion. Five treatments were compared in each group, and the clinical outcomes were evaluated during follow-ups. RESULTS: A 5-aminolevulinic acid-mediated photodynamic therapy (ALA-PDT) is preferred if the maximum lesion diameter is <0.5 cm and an ALA-PDT plus cryotherapy treatment is preferred for lesions 0.5-2.0 cm. For lesions >2.0- 4.0 cm, an ALA-PDT retreatment (after cryotherapy or CO2 laser treatment) should be the first choice. CONCLUSIONS: The treatments for CA should be chosen according to the maximum diameter of each patient's lesion.

The Role of EGFR/ERK/ELK-1 MAP Kinase Pathway in the Underlying Damage to Diabetic Rat Skin.

BACKGROUND: Diabetes mellitus (DM) is a highly prevalent disease. Atrophy and spontaneous ulcers are the most common cutaneous manifestation of diabetic dermopathy (DD). Before spontaneous ulcers, we believe there is an underlying damage stage although the mechanism is unknown. AIMS: To explore the expression of extracellular signal-regulated kinase1/2 (ERK1/2), its correlated upstream protein epidermal growth factor receptor (EGFR) and its downstream transcription factor E twenty-six (ETS)-like 1(ELK-1)in the damage of the diabetic rat skin, and to explore the role of ERK1/2 on the recessive damage to diabetic rat skin. MATERIALS AND METHODS: Eighty Sprague-Dawley (SD) rats weighing 260-300 g were randomly divided into control and streptozotocin (STZ)-induced diabetes groups. After 0.5, 2, 4, and 8 weeks, the shaved skin specimens from the back of rats in both groups were collected to observe the histological characteristics of the skin, to measure the thickness of the epidermis and the dermis, and to observe the ultrastructure. Immunohistochemistry (IHC) and Western blot techniques were used to detect the expression and activation of ERK1/2, EGFR, ELK-1 in the skin of the rats. RESULTS: There are ultrastructural changes in the DM skin. With the continuance of the diabetes course, the thicknesses of the epidermis and dermis decreased, and the expression of phospho-ERK1/2 (P-ERK1/2), EGFR, and ELK-1 was decreased gradually in the back skin of the diabetes rats. It was significantly lower in 4 and 8 week DM than that of the normal control (P < 0.05). The expression of P-EGFR and P-ERK1/2 in the back skin of the diabetes rats was positively correlated (r = 0.572 P < 0.05), and the positive correlation was also obtained between P-ERK1/2 and P-ELK-1 (r = 0.715, P < 0.05). CONCLUSION: THE PHENOMENON OF RECESSIVE DAMAGE EXISTS IN THE SKIN OF DIABETES RATS, WHICH PROBABLY MAY RELATE TO THE WEAKNESS OF THE SIGNAL TRANSDUCTION: P-EGFR → ERK1/2 → ELK-1.