Expression of miR-145 and miR-18b in Peripheral Blood Samples of Head and Neck Cancer Patients.

Head and neck squamous cell carcinomas (HNSCC) is one of the most prevalent type of cancer known in Indian population. Studies are needed to identify the early biomarkers for HNSCC. MicroRNAs (miRNAs) are non-coding RNA molecules, expression of which can be used as biomarker for early diagnosis of HNSCC. For miRNA profiling total RNA, which also contained small RNAs were isolated from ten HNSCC tissue samples and adjacent control. Purity and concentration of eluted RNA was assessed using the NanoDrop1000® spectrophotometer, Reverse Transcription reaction was carried out with megaplex RT primers of pool A and pool B and the expression of selected miRNAs (miR-143/145 and miR-18a/b) was measured using TaqMan primers specific for mature miRNAs. Our study showed dramatic downregulation in expression of two miRNAs, miR-18b and miR-145 in blood samples of HNSCC patients, which are inhibitor of tumorigenesis and can be targeted as biomarker of HNSCC pathogenesis therefore developing avenues for miRNA role in prognosis and therapeutics. Supplementary Information: The online version contains supplementary material available at 10.1007/s12291-023-01119-2.

Short-course radiotherapy with consolidation chemotherapy versus conventionally fractionated long-course chemoradiotherapy for locally advanced rectal cancer: randomized clinical trial.

BACKGROUND: The trial hypothesis was that, in a resource-constrained situation, short-course radiotherapy would improve treatment compliance compared with conventional chemoradiotherapy for locally advanced rectal cancer, without compromising oncological outcomes. METHODS: In this open-label RCT, patients with cT3, cT4 or node-positive non-metastatic rectal cancer were allocated randomly to 5 × 5 Gy radiotherapy and two cycles of XELOX (arm A) or chemoradiotherapy with concurrent capecitabine (arm B), followed by total mesorectal excision in both arms. All patients received a further six cycles of adjuvant chemotherapy with the XELOX regimen. The primary endpoint was treatment compliance, defined as the ability to complete planned treatment, including neoadjuvant radiochemotherapy, surgery, and adjuvant chemotherapy to a dose of six cycles. RESULTS: Of 162 allocated patients, 140 were eligible for analysis: 69 in arm A and 71 in arm B. Compliance with planned treatment (primary endpoint) was greater in arm A (63 versus 41 per cent; P = 0.005). The incidence of acute toxicities of neoadjuvant therapy was similar (haematological: 28 versus 32 per cent, P = 0.533; gastrointestinal: 14 versus 21 per cent, P = 0.305; grade III-IV: 2 versus 4 per cent, P = 1.000). Delays in radiotherapy were less common in arm A (9 versus 45 per cent; P < 0.001), and overall times for completion of neoadjuvant treatment were shorter (P < 0.001). The rates of R0 resection (87 versus 90 per cent; P = 0.554), sphincter preservation (32 versus 35 per cent; P = 0.708), pathological complete response (12 versus 10 per cent; P = 0.740), and overall tumour downstaging (75 versus 75 per cent; P = 0.920) were similar. Downstaging of the primary tumour (ypT) was more common in arm A (P = 0.044). There was no difference in postoperative complications between trial arms (P = 0.838). CONCLUSION: Reduced treatment delays and a higher rate of compliance were observed with treatment for short-course radiotherapy with consolidation chemotherapy, with no difference in early oncological surgical outcomes. In time- and resource-constrained rectal cancer units in developing countries, short-course radiotherapy should be the standard of care.

Feasibility, efficiency & effectiveness of pooled sample testing strategy (pooled NAAT) for molecular testing of COVID-19.

BACKGROUND & OBJECTIVES: During the current COVID-19 pandemic, a large number of clinical samples were tested by real-time PCR. Pooling the clinical samples before testing can be a good cost-saving and rapid alternative for screening large populations. The aim of this study was to compare the performance characteristics, feasibility and effectiveness of pooling nasal swab and throat swab samples for screening and diagnosis of SARS-CoV-2. METHODS: The pool testing was applied on a set of samples coming from low COVID-19 positivity areas. A total of 2410 samples were tested in pools of five samples each. A total of five pools of five samples each were generated and tested for E gene. RESULTS: Of the total of 482 pools (2410 samples) 24 pools flagged positive. Later on pool de-convolution, a total of 26 samples were detected as positive for COVID-19, leading to positivity of about one per cent in the test population. For the diagnosis of individual samples, the pooling strategies resulted in cost savings of 75 per cent (5 samples per pool). INTERPRETATION & CONCLUSIONS: It was observed that testing samples for COVID-19 by reverse transcription (RT)- PCR after pooling could be a cost-effective method which would save both in manpower and cost especially for resource-poor countries and at a time when test kits were short in supply.

A study on local expression of NF-κB, CCL2 and their involvement in intratumoral macrophage infiltration in breast cancer.

NF-κB has been implicated in mechanisms promoting inflammation in tumor microenvironment leading to breast cancer metastasis. Owing to critical role of CCL2 during metastasis, particularly in its capacity to act as a chemoattractant for macrophages and their precursors i.e monocytes, we decided to explore if pro-metastatic function of NF-κB could be attributable to CCL2 and/or macrophage infiltration. Through our study we provide experimental and clinical evidence in support of co-ordinated expression of chemokines CCL2, NF-κB and intratumoral macrophage content particularly with respect to breast cancer, with an additional evidence of these three variables being key determinant for poor prognosis and diminished survival amongst breast cancer patients both independently as well in a coordinated manner. The mean fold increase in mRNA expression level of NF-κB and CCL2 indicated that it was over expressed 13.57 and 13.18 fold respectively in tumor tissue as compared to adjacent normal tissue. Among these Immunohistochemistry expression of CD68 marker showed that 62 patients (66.7%) had low/moderate CD68 expression while 31 patients (33.3%) had strong expression. All three variables viz.NF-κB, CCL2 and CD68 showed significant (p<0.05 or p<0.01 or p<0.001) respectively associations with both clinicopathological (except CD68 with stage) and hormone receptors (ER, PR and Her2/neu) and their co-expressions indicating these as predictors of breast cancer. In this study we decipher the possible molecular mechanism by way of which NF-κB may promote breast cancer metastasis. Our study has clinical relevance as it establishes significance of these three variables as potential predictive markers to be employed in breast cancer.

Evaluation of urinary XIAP as a diagnostic biomarker of carcinoma of urinary bladder.

Early diagnosis of carcinoma of the bladder remains a challenge. Urine cytology, as an adjunct to cystoscopy, is less sensitive for low-grade tumors. X-linked inhibitor of apoptosis (XIAP), a negative regulator of apoptotic stimuli and a member of apoptosis family, is frequently activated in bladder carcinoma. Our aim is to investigate the significance of urinary XIAP for the noninvasive diagnosis of transitional cell carcinoma (TCC) of the urinary bladder. We examined urinary XIAP expression in a bladder cancer cell line (T24) and in urine of 28 healthy individuals, 46 patients of nonmalignant disorders, and 117 cases (69 primary and 48 recurrent cases) of histologically proven TCC prior to transurethral resection, by using real-time PCR, and compared it with voided urinary cytology (VUC). XIAP expression was found in T24 cell line and also was found to be significantly higher in the cancer group as compared to the controls (p < 0.001). XIAP messenger RNA (mRNA) expression showed a significant (p < 0.05) association with stage and grade (p < 0.05). XIAP shows the sensitivity of 82.91 % and specificity of 78.38 % (p < 0.001), whereas urine cytology had sensitivity of 66.67 % and specificity of 95.95 % for TCC cases. The combination of XIAP and VUC had better sensitivity (98.2 %) and specificity (92.6 %) than they showed individually (p < 0.001). XIAP mRNA expression did not significantly correlated with the patient's age, sex, and smoking (p > 0.05). Urinary XIAP can be used as a noninvasive diagnostic biomarker for bladder TCC in adjunct to cytology mainly for low-grade non-muscle-invasive tumors.

Appraisal of diagnostic ability of UCA1 as a biomarker of carcinoma of the urinary bladder.

Initial diagnosis of carcinoma of the urinary bladder remains to be a challenge. Urine cytology, as an adjunct to cystoscopy, is less sensitive for low-grade tumors. Urothelial cancer associated 1 (UCA1) is a novel non-coding RNA gene, which plays a pivotal role in bladder cancer progression. Our aim is to investigate the significance of urinary UCA1 for the non-invasive diagnosis of transitional cell carcinoma (TCC) of the urinary bladder. We examined UCA1 expression in a bladder cancer cell line (T24) and in urine of 28 healthy individuals, 46 patients of non-malignant disorders, and 117 cases (69 primary and 48 recurrent cases) of histologically proven TCC prior to transurethral resection by using real-time PCR and compared it with voided urinary cytology. UCA1 expression was found in T24 cell line and also found to be significantly higher in the cancer group as compared to the controls (p<0.001). UCA1 messenger RNA (mRNA) expression showed a significant (p<0.05) association with stage and grade (p<0.05). UCA1 showed a sensitivity of 79.49% and a specificity of 79.73% (p<0.001), whereas urine cytology had a sensitivity of 66.67% and a specificity of 95.95% for TCC cases. Higher expression of UCA1 was associated with high grade (G2-G3, sensitivity=84.09%) (p<0.001). UCA1 mRNA expression did not significantly correlate with the patient's age, sex, and smoking habit (p>0.05). UCA1 can be used as a non-invasive diagnostic biomarker for TCC bladder as an adjunct to cytology in the early diagnosis of primary urinary bladder cancer.

Cyclooxygenase-2 inhibition attenuates hypoxic cancer cells induced m2-polarization of macrophages.

Tumor-associated macrophages (TAMs), represent a major subpopulation of tumor infiltrating immune cells. These alternatively activated M2-polarized macrophages are well known for their pro-tumor functions. Owing to their established role in potentiating tumor-neovasculogenesis and metastasis, TAMs have emerged as promising target for anti-cancer immunotherapy. One of the key TAMs related phenomenon that is amenable to therapeutic intervention is their phenotype switching into alternatively activated M2-polarized macrophages. Hindering macrophage polarization towards a pro-tumor M2 phenotype, or better still reprogramming the M2 like TAMs towards M1 subtype is being considered a beneficial anti-cancer strategy. Hypoxic tumor milieu has been proposed as one of the most plausible factor governing M2-polarization of macrophages. We recently demonstrated that hypoxic tumor cells imparted a pro­angiogenic M2 skewed phenotype to macrophages. Furthermore, sizeable body of data indicates for participation of cyclooxygenase-2 (COX-2) in macrophage polarization. Concordantly, inhibition of COX-2 is associated with impaired macrophage polarization. Prompted by this in the current study we decided to explore if inhibition of COX-2 activity via chemical inhibitors may prevent hypoxic cancer cell induced M2-polarization of macrophages. We observed that treatment with Flunixin meglumine, an established preferential inhibitor of COX-2 activity markedly inhibited hypoxic cancer cell induced of M2-polarization of macrophages thereby indicating for usage of COX-2 inhibition as possible anti-cancer treatment modality.

Assessment of Uncertainty in Volume Estimation of Non-Static Target: A Phantom Study using Racemosa Wood.

INTRODUCTION: CT information of the target undergoing motion/movement during its scanning has been questioned by many researchers for its preciseness as well as accuracy. The present study was taken with aim to validate the racimosa wood as lung equivalent and to assess the uncertainty in volume estimation during virtual simulation of non-static target of known dimension such as in lung cancer radiotherapy. MATERIALS AND METHODS: The racemosa wood was validated as lung equivalent material with the help of two methods. Wood insert with tumor model was put into the hollow cylinder space of dimension 6.4 cm in diameter provided in CIRS phantom. First CT image of rest position was taken and given name "No Movement". Subsequently the tumor was shifted +/-5mm, +/-15mm and +/-25 mm with respect to "Rest Position". CT images of the CIRS phantom containing tumor in wood cylinder were acquired after each movement given to wood cylinder. RESULTS: The relative electron density of racemosa wood corresponding to HU value -724 was found to be 0.275 gm/cm3. The true volume of the target was 7.8.cm3 however variation up to 9.5 cm3 was observed in CT produced volume of the target over the range of different movements. DISCUSSION: The racemosa wood was found to be having range of density (- 850 HU to - 400 HU) similar to real human lung density variation. Various studies have been performed using uniform density lung structures in their experimental setups to assess the accuracy in lung cancer radiation delivery. However, in the present work approximately real clinical setting was reproduced by putting the wood cylinder with density variation from 0.2 gm/cm3- 4.5 gm/cm3in hollow space provided in one lung structure the phantom used in this study. CONCLUSIONS: The racemosa wood was found to be lung equivalent which is available locally and cost-effective as well. Overestimation in the target volume (by CT imaging) showed a trend of increase with 3 directional movement amplitudes. The results of this study can be utilised in lung cancer radiotherapy as the same were derived from setup having clinical settings in terms of lung density variation, shape, compositions of the phantom maximally as found during the real patient radiotherapy.

Centchroman inhibits proliferation of head and neck cancer cells through the modulation of PI3K/mTOR pathway.

Centchroman (CC; 67/20; INN: Ormeloxifene) is a non-steroidal antiestrogen extensively used as a female contraceptive in India. In the present study, we report the anti-proliferative effect of CC in head and neck squamous cell carcinoma (HNSCC) cells. CC inhibited cell proliferation in a dose dependent manner at 24 h of treatment. Further studies showed that CC treatment induced apoptosis, inhibited Akt/mTOR and signal transducers and activators of transcription protein 3 (STAT3) signaling, altered proteins associated with cell cycle regulation and DNA damage and inhibited colony forming efficiency of HNSCC cells. In addition, CC displayed anti-proliferative activity against a variety of non-HNSCC cell lines of diverse origin. The ability of CC to serve as a dual-inhibitor of Akt/mTOR and STAT3 signaling warrants further studies into its role as a therapeutic strategy against HNSCC.

Tumefactive demyelinating lesion: experience with two unusual patients.

Tumefactive demyelinating lesion, a variant of multiple sclerosis, is a solitary large demyelinating lesion, which mimics cerebral neoplasm. Distinguishing tumefactive lesions from other etiologies of intracranial space-occupying lesions is essential to avoid inadvertent surgical or toxic chemotherapeutic interventions. We report two unusual cases of tumefactive demyelinating lesion. The first patient presented with recurrent right focal tonic-clonic seizures with secondary generalization of three-month duration. Her neurological examination was normal. Another patient presented with right homonymous hemianopia. In this patient, the diagnosis was established after biopsy of the lesion, which revealed perivascular lymphocytic infiltrate and aggregates of foam cells in white matter with relatively uninvolved grey matter, suggestive of tumefactive demyelinating lesion. Administration of intravenous methylprednisolone resulted in rapid clinical improvement in both the patients and the brain lesion decreased in size. Both, epilepsy and homonymous hemianopia, are unusual manifestations of tumefactive demyelinating lesions. In our cases, cerebral tumors were initial diagnoses. Presence of an open ring or incomplete ring lesions and other magnetic resonance characteristics helped in differentiating demyelinating lesions from other neoplastic and infective diseases of the brain. Differential diagnosis of tumefactive demyelinating lesions, at times, may prove to be a challenging task for the treating physician.

Assessment of free radical-mediated damage in head and neck squamous cell carcinoma patients and after treatment with radiotherapy.

Reactive oxygen species (ROS) produced as a part of cellular metabolism can interact with biological macromolecules such as DNA, proteins and lipids and interfere with their normal functions, leading to the loss of cellular viability. ROS have been implicated in many pathophysiological conditions including cancer. In the present study, the damage caused by ROS and the effect of radiation in head and neck squamous cell carcinoma (HNSCC) patients were assessed in the erythrocytes by analyzing the superoxide dismutase (SOD) and catalase (CAT) activities, and levels of total thiols (T-SH) and malondialdehyde (MDA, a marker for lipid peroxidation). Blood samples were collected before the start of treatment and after the completion of radiotherapy. Both SOD and CAT activities were decreased in untreated patients, but elevated in patients after treatment. The T-SH levels were also depleted in untreated HNSCC patients, but elevated non-significantly after radiation therapy (p>0.05). The levels of MDA showed a significant increase in both untreated patients and after radiation therapy when compared with normal subjects (p<0.05). Thus, the present study indicated that the free radical-mediated damage was aggravated in untreated HNSCC patients, but the levels of antioxidants returned to baseline or nearly so after the treatment with radiation therapy.

Prospective evaluation of XRCC-1 Arg194Trp polymorphism as bio-predictor for clinical outcome in locally advanced laryngeal cancer undergoing cisplatin-based chemoradiation.

BACKGROUND: To determine X-ray repair cross-complementing 1 gene (XRCC-1) Arg194Trp polymorphism as bio-predictor for clinical outcome in advanced laryngeal squamous cell carcinoma undergoing cisplatin-based chemoradiation (CRT). METHODS: A total of 150 patients were enrolled in this prospective study. XRCC-1 Arg194Trp genotyping categorized patients as wild (C/C) and polymorphic (C/T or T/T). The primary endpoint was to assess acute radiation-induced toxicity (ARIT). RESULTS: A significant correlation of skin (P- .04) and oral mucosal ARIT (P- .01) was noticed in the XRCC-1 polymorphic variant. A higher treatment response was noted in the polymorphic variant, and it shows a trend toward significance (P- .08). With 33 months of median follow-up, 2-year progression-free survival (PFS) and overall survival (OS) of wild vs polymorphic variant were 34.6% vs 46.9% (P- .066) and 50.6% vs 62.2% (P- .12). CONCLUSION: XRCC-1 polymorphic variants have significantly higher grade of >2 ARIT and may have improved trend for treatment response and PFS.

Alteration of the Risk of Oral Pre-Cancer and Cancer in North India Population by CYP1A1 Polymorphism Genotypes and Haplotype

Background: The aim of this study was to evaluate any association between CYP1A1 (T6235C and C4887A, A4889G) gene polymorphisms and the risk of oral pre-cancer and cancer. Methods: In the present study, 250 patients with oral pre-cancer and/or cancer and 250 healthy controls were genotyped for CYP1A1 T6235C, C4887A and A4889G polymorphisms by the PCR-RFLP method. Results: None of the CYP1A1 polymorphisms were associated with the risk of either oral cancer or pre cancer. Nor were any links with clinical parameters of oral cancer found. However, among the consumers of areca nut/pan masala the TC, CA and AG genotypes respectively for the CYP1A1 T6235C,C4887Aand A4889G polymorphisms were significantly more frequent in controls compared to cases (p values for cases vs. controls of 0.0032, 0.0019 and 0.0009, respectively). Similarly, compared to the haplotype TCA, TAG constituted by CYP1A1 T6235C and C4887A and A4889G was more common in controls (6.88%) than in cases (4.07%). Conclusion: Our results suggest that genotypes regarding CYP1A1 polymorphisms may modulate the risk of oral cancer and pre-cancer among the areca nut/pan masala consumers. The haplotype may also exert an influence in our north Indian population.

Evaluation of Lung Density and Its Dosimetric Impact on Lung Cancer Radiotherapy: A Simulation Study.

BACKGROUND: The dosimetric parameters required in lung cancer radiation therapy are taken from a homogeneous water phantom; however, during treatment, the expected results are being affected because of its inhomogeneity. Therefore, it becomes necessary to quantify these deviations. OBJECTIVE: The present study has been undertaken to find out inter- and intra- lung density variations and its dosimetric impact on lung cancer radiotherapy using Monte Carlo code FLUKA and PBC algorithms. MATERIAL AND METHODS: Density of 100 lungs was recorded from their CT images along with age. Then, after PDD calculated by FLUKA MC Code and PBC algorithm for virtual phantom having density 0.2 gm/cm3 and 0.4 gm/cm3 (density range obtained from CT images of 100 lungs) using Co-60 10 x10 cm2 beams were compared. RESULTS: Average left and right lung densities were 0.275±0.387 and 0.270±0.383 respectively. The deviation in PBC calculated PDD were (+)216%, (+91%), (+)45%, (+)26.88%, (+)14%, (-)1%, (+)2%, (-)0.4%, (-)1%, (+)1%, (+)4%, (+)4.5% for 0.4 gm/cm3 and (+)311%, (+)177%, (+)118%, (+)90.95%, (+)72.23%, (+)55.83% ,(+)38.85%, (+)28.80%, (+)21.79%, (+)15.95%, (+)1.67%, (-) 2.13%, (+)1.27%, (+)0.35%, (-)1.79%, (-)2.75% for 0.2 gm/cm3 density mediums at depths of 1mm, 2mm, 3mm, 4mm, 5mm, 6 mm, 7 mm, 8mm, 9mm,10mm, 15mm, 30mm, 40mm, 50mm, 80mm and 100 mm, respectively. CONCLUSION: Large variations in inter- and intra- lung density were recorded. PBC overestimated the dose at air/lung interface as well as inside lung. The results of Monte Carlo simulation can be used to assess the performance of other treatment planning systems used in lung cancer radiotherapy.

Evaluation of Volumetric Doses of Organs at Risk in Carcinoma Cervix Patients with HDR Intracavitary Brachytherapy and Comparison of CT-based and Conventional Plans.

BACKGROUND: Brachytherapy treatment planning in cervix carcinoma patients using two dimensional (2D) orthogonal images provides only point dose estimates while CT-based planning provides volumetric dose assessment helping in understanding the correlation between morbidity and the dose to organs at risk (OARs) and treatment volume. OBJECTIVE: Aim of present study is to compare International Commission on Radiation Units and Measurements Report 38 (ICRU 38) reference point doses to OARs with volumetric doses using 2D images and CT images in patients with cervical cancer. MATERIAL AND METHODS: In this prospective study, 20 patients with cervical cancer stages (IIB-IIIB) were planned for a brachytherapy dose of 7Gy per fraction for three fractions using 2D image-based treatment plan and CT-based plan. ICRU 38 points for bladder and rectum were identified on both 2D image-based plan and CT-based plan and doses (DICRU) at these points were compared to the minimum dose to 2cc volume (D2cc) of bladder and rectum receiving the highest dose. RESULTS: D2cc bladder dose was 1.60 (±0.67) times more than DICRUb bladder dose whereas D2cc rectum dose was 1.13±0.40 times DICRUr. Significant difference was found between DICRUb and D2cc dose for bladder (p=.0.016) while no significant difference was seen between DICRUr and D2cc dose for rectum (p=0.964). CONCLUSION: The study suggests that ICRU 38 point doses are not the true representation of maximum doses to OARs. CT-based treatment planning is more a reliable tool for OAR dose assessment than the conventional 2D radiograph-based plan.

Validation of the Gel & Wax Boluses and Comparison of their Dosimetric Performance with Virtual Bolus.

BACKGROUND: In general, radiotherapy treatment planning is performed using the virtual bolus. It is necessary to investigate physical bolus in comparison to virtual one. OBJECTIVES: In the present study, first, radiological properties of superflab Gel bolus and Paraffin wax bolus was investigated in terms of their relative electron density. Then, dosimetric performance of both the bolus (i.e. Gel and Parafin wax) was compared with Virtual bolus. MATERIAL AND METHODS: In This experimental study, the radiological property of Wax and Gel boluses was investigated using two methods. In one, the relative electron density of both the Gel and Wax boluses was calculated by measuring their linear attenuation coefficient where in another method relative electron density was calculated by recording their CT No directly from their CT scan. Later CT scan of solid water slab phantom (dimension 30x30x15 cm3), with physical boluses (i.e. Gel and Wax bolus) of appropriate thicknesses required to deliver a dose of 200 cGy at Dmax using 4 MV, 6 MV and 15 MV photon beams, was taken. These CT data sets were retrieved to TPS. A plan was done to deliver a dose of 200 cGy at Dmax using Single 4 MV, 6 MV and 15 MV photon beams. Dose at depths Dmax, 1 cm, 2 cm, 3 cm, 4 cm and 5 cm was recorded. Using this similar method, doses at depths viz Dmax, 1 cm, 2 cm, 3 cm, 4 cm and 5 cm was recorded for the Gel and Wax boluses. The differences in dose of gel and wax bolus from virtual bolus were recorded for comparison of their dosimetric performance. RESULTS: The measured (calculated) relative electron density of wax and Gel bolus was found to be 0.958 (0.926) and 0.923 (0.907), respectively. Variation in dosimetric performance of Gel and Wax with reference to Virtual bolus was studied. However, on average, Gel bolus was more consistent with virtual bolus. CONCLUSION: To avoid any dose difference between, delivered (using physical bolus) and planned (using virtual bolus), the physical boluses should be investigated for their dosimetric performance in comparison to virtual bolus. The results obtained and methodology used in this study can be applied in routine radiotherapy practices.

Tissue and serum expression of TGM-3 may be prognostic marker in patients of oral squamous cell carcinoma undergoing chemo-radiotherapy.

Radioresistance is one of the main determinants of treatment outcome in oral squamous cell carcinoma (OSCC), but its prediction is difficult. Several authors aimed to establish radioresistant OSCC cell lines to identify genes with altered expression in response to radioresistance. The development of OSCC is a multistep carcinogenic process that includes activation of several oncogenes and inactivation of tumour suppressor genes. TGM-3 is a tumour suppressor gene and contributes to carcinogenesis process. The aim of this study was to estimate serum and tissue expression of TGM-3 and its correlation with clinico-pathological factors and overall survival in patients of OSCC undergoing chemo-radiotherapy. Tissue expression was observed in formalin fixed tissue biopsies of 96 cases of OSCC and 32 healthy controls were subjected to immunohistochemistry (IHC) by using antibody against TGM-3 and serum level was estimated by ELISA method. mRNA expression was determined by using Real-Time PCR. Patients were followed for 2 year for chemo radiotherapy response. In OSCC, 76.70% cases and in controls 90.62% were positive for TGM-3 IHC expression. TGM-3 expression was cytoplasmic and nuclear staining expressed in keratinized layer, stratum granulosum and stratum spinosum in controls and tumour cells. Mean serum TGM-3 in pre chemo-radiotherapy OSCC cases were 1304.83±573.55, post chemo-radiotherapy samples were 1530.64±669.33 and controls were 1869.16±1377.36, but difference was significant in pre chemo-radiotherapy samples as compared to controls (p<0.018). This finding was also confirmed by real- time PCR analysis in which down regulation (-7.92 fold change) of TGM-3 in OSCC as compared to controls. TGM-3 expression was significantly associated with response to chemo-radiotherapy treatment (p<0.007) and overall survival (p<0.015). Patents having higher level of TGM-3 expression have good response to chemo-radiotherapy and also have better overall survival. TGM-3 may serve as a candidate biomarker for responsiveness to chemo-radiotherapy treatment in OSCC patients.

Correction: Tissue and serum expression of TGM-3 may be prognostic marker in patients of oral squamous cell carcinoma undergoing chemo-radiotherapy.

[This corrects the article DOI: 10.1371/journal.pone.0199665.].